Medical Biochemistry and Molecular Basis of Medical Genetics Metabolism Overview 1.

Define the terms metabolism, metabolites and metabolic pathway. --metabolism sum of all enzyme-catalyzed reactions that occur in a living organism. Dynamic, coordinated, highly regulated. !"!#$%. --metabolites small mol. involved in the synthesis & degradation of biopolymers & the interconversion of chemical compounds 'o(idation of glucose, fats, etc. to $)*+, e(. fatty acids, polypeptides, etc. -he *-$ acetyl group is -.% central intermediate in most !-/ production pathways. --metabolic pathway series of coupled r(s in which prod. of 1 r( acts as the substrate for the ne(t r( w0 regulation of the flow of metabolites. *. $ompare and contrast catabolism and anabolism in terms of what they are and whether they wor1 to use energy or store energy. --catabolism breaking down biomolecules in order to produce energy 'via oxidation+ & the building bloc1s for other synthesis. /athways coverge on only a few products, all of which enter the -$! cycle. RELEA E energy to 2-)3%. --anabolism building up. Biosynthesis of more comple( molecules from small precursors in reductive pathways 'i.e. ! E energy+. !nabolic pathways are divergent starting w0 a few metabolites & producing many diff. molecules 'acetyl $o! can be built upon to ma1e most molecules, etc+. 4. %(plain the central role of nucleotide triphosphates, reduced coenzymes and acetyl coenzyme ! in metabolism. --lead to production of !-/ by leading to production of reduced co-enzymes. --energy released by catabolism is conserved in these molecules 5. %(plain the concept of balance in metabolic pathways. --metab. pathways designed to meet specific cellular needs. $an be source of metabolites as needed by other pathways, can use metabolites as needed from other pathways. 6etab. series of coupled r(s. 'prod. of 1 r( substrate for ne(t r(+ 7. Define and e(plain substrate cycles in metabolic pathways. 8nclude a consideration of futile cycles and how metabolic pathways prevent this. --paired cycles of reactions in which one cycle proceeds one direction and the other cycle proceeds bac1, when the degradative and synthetic pathway for a biomolecule uses the same pathway 'w0 some 1ey differences+. 9ycolysis and gluconeogenesis. %a. pathway is controlled by sep. reg. ezs which are reciprocally regulated to prevent futile cycles 'endless little cycles of going bac1 and forth in one small intermediate r(.+ :. Describe how compartmentalization of metabolic pathways allows for efficient regulation of the pathways. --comp. occurs w0in cells by restricting pathways to particular subcellular organelles and w0in multicellular organisms by specialization of tissues. ;. %(plain the flow of metabolites during the fed state. 8n particular, e(plain the goal of metabolite utilization and the specific processes that occur in different tissues 'slide *5+.

--fuels consumed are o(idized to meet immediate needs. %(cess fuel is stored as triacylglycerol and glycogen. --once fuels enter the intestines, carbs are bro1en down to glucose, fats are prepac1ed into chylomicrons, and amino acids go directly to the tissues to be used in the manufacture of proteins and the -$! cycle. 9lucose triggers an in insulin and a in glucagon in the blood. 8t also travels to the liver where it is bro1en down into glycogen and0or triglycerides 'destined for <"D"+ & acetyl $o! ' -$! cycle+. 9lucose also travels to brainacetyl $o!-$!, to the 3 $spyruvatelactate, to muscle for acetyl $o!-$! or glycogen storage, or to adipose tissue as triglycerides. )verall, glucose = insulin & glucagon. >. "ist the caloric content of carbohydrates, fats, protein and alcohol. --carbohydrates? 5 1cal0g --protein? 5 1cal0g --fats? @ 1cal0g --alcohol? ; 1cal0g @. $alculate calories consumed given specific Auantities of carbohydrate, fat, protein and0or alcohol. -- carb. grams ( 51cal0g B fat g ( @1cal0g B protein g ( 51cal0g B alcohol g ( ;1cal.g 1C. "ist and e(plain our dietary reAuirements for carbohydrate, fats, protein, vitamins and minerals and water. --no carbs are essential, we can synthesize any we need. --essential fats? -linoleic acid & -lenolenic acid, eicosapentaienoic acid '%/!+ and docosahe(anoic acid 'D.!+, from fish oils 'omega 4 fats+ --protein? @ essential a.a.s 'D8"-<EF6.+ 'arg conditionally essential+, nitrogen balance reAuired. --certain vitamins and minerals are essential --water essential 11. %(plain how consumed food is bro1en down and absorbed. --food bro1en down into smallest units by series of enzymes in mouth, stomach, sm. intest, carbsmonosaccharides, proteinssingle a.a.s, fats chylomicrons 1*. %(plain the relationship between glucose and insulin, and insulin and glucagons. --glucose in blood stimulates prod. of insulin by pancreas. --insulin stimulates cells to ta1e up glucose --insulin causes glucagon levels to decrease 'glucagon signals fasting state+ 14. Describe the fate of glucose after a meal in the following tissues? muscle, brain, liver, red blood cells, adipose tissues. --muscle uses blood glucose for energy & to maintain glycogen stores in muscle --brain highly dependent on glucose as the maGor energy source, fatty acids canHt pass blood-brain barrier, glucose supply tightly regulated to ensure enough glucose is provided. --liver some glucose immed. metab. to form !-/ to meet energy needs. remainder converted to storage as glycogen & triaclyglycerol --3 $s glucose only source of energy. no mitochondria for utilizing other metabolites, blood levels of glucose tightly regulated to ensure 3 $ 'and other tissues+ have enough glucose. 3 $ survival depends on this directly. --adipose tissue insulin greatly stimulates glucose upta1e into adipose tissue. glucose is o(id. for energy & provides glycerol moiety for ma1ing -!9s for storage 15. Describe the fate of lipoproteins and amino acids after a meal. 2

--lipoproteins chylomicrons & <"D" produced by liver after ingesting fats transport -9 & cholesterol in blood, <"D" bind to adipose cells, -9 degraded to free F!, F!s transported into adipose cells, reformed to -!9s using glycerol formed from blood glucose & stored as fat droplets in cells. 3emnants of <"D" cleared by liver, or used to ma1e low density lipoproteins '"D"s+. --amino acids a.a. from dietary protein ta1en up by various tissues, most used to synthesize new protein, some o(idized to yield energy or to ma1e important metabolites I!-/, hormones, neurotransmitters, heme, etc, a.a.s produced by protein degradation enter same pool as dietary a.a.s, used for new protein synthesis, muscle is maGor user of a.a.s to ma1e new protein b0c of maGor mass of muscle tissue in body. 17. %(plain the flow of metabolites during the fasting state 'slide 4C+. 8n particular, e(plain the goal of metabolite utilization and the specific processes that occur in different tissues. --body uses fuel stores to provide energy for tissues, esp. the brain. 2trategies to meet energy needs in short term 'fasting+ diff. from long term 'starvation+. --glucose = glucagon and insulin in blood --adipose tissue releases -9 which are converted to glycerol for use in the liver or F!s for use in muscle as acetyl $o! -$! or for use in liver as acetyl $o!1etone bodies 'bac1 to acetyl $o! in muscle+. 9lycerol in liver is converted to glucose for use in the brain and 3 $s and urea waste is funneled thru 1idneys --protein from muscle tissue also bro1en down to glucose in liver. 1:. "ist the body stores of carbohydrate, fat and protein. --carbs - stored as glycogen 'glucose storage in liver, muscle and other cells+, produces 51cal0gm when utilized --fat stored in adipose tissue as triacylglycerol at @1cal0gm utilized --protein contained in s1eletal muscle which provides short-term a.a.s J 51cal0g 1;. Describe the metabolic changes that occur during fasting in tissues, and especially adipose tissue and liver. --blood glucose levels pea1 K 1hr. after eating. * hrs. after eating = fasting range --glucose levels signal pancreas to stop prod. insulin & insulin --liver responds to insulin by glycogen degradation to release glucose into blood --1* hrs. after eating, body is in basal 'post-absorptive+ state. 2erum insulin levels are , liver glycogen stores are . 9lucagon level is rising & gluconeogenesis 'util. of lactate, glycerol, a.a.s to made glucose+ is starting to supply blood glucose. L- most F!s canHt provide $ for gluconeogenesis, only the glycerols of -9 can be used. F!s o(idized directly to yield energy. --adipose tissue F!s provide maGor source of fuel during fasting. F! o(idized to acetyl $o! !-/ prod. 9lycerol bac1bone used for gluconeogenesis. --liver most F! ta1en up used to produce D%-)#% )D8%2, which can be used by some tissues 'muscle, 1idney+ 1>. %(plain the flow of metabolites during a long fast 'slide 4:+. 8n particular, e(plain 1ey changes in metabolite production and utilization 'i.e., 1etone bodies+ and the specific processes that occur in different tissues 'esp. muscle, liver, brain adipose tissue+. --glucose levels , insulin levels , glucagon levels 3

--very little glucose goes to brain in long fast, so 1etone bodies used instead. ---9 from adipose glycerol to liverglucose to brain, 3 $s, -9 F!s to acetyl $o! in muscle and liver. "iver acetyl $o!1etone bodies for use in brain. --protein in muscles a.a.s to liver glucose, waste in urea to 1idneys --2ummary? emphasis shifts away from prod. glucose as maGor fuel which use of a.a. from protein degradation, sparing muscle mass, muscle use of 1etone bodies and uses F! o(id. as primary source of fuel. rian starts to ta1e up & utilize 1etone bodies to ma1e acetyl $o!, a.a.s only used for gluconeogenesis to provide glucose to blood for 3 $s. brain uses glucose, more avail. for 3 $s & less needed to be made from a.a. degradation. a.a. used for gluconeogenesis, e(cess nitrogen so ed urea prod. 1@. Define 63 and 68. -- 63 = basal metabolic rate = energy reAuired to sustain life -- 68 = body mass inde( = method for determining if weight is in healthy range *C. %(plain the relationship between daily energy e(penditure and diet. --energy balance = consuming amnt of food eAual to daily energy e(penditure --consume more food = store more fuel = gain more weight --consume less food = use stores = lose weight Lectures "# and "$% Bioenergetics 3eference? 2mith, 6ar1s and "ieberman $hapter 1@. 1. Define the term thermodynamics and e(plain its relevance to biological systems. --collection of laws & principles describing the flows & interchanges of heat, energy, and matter in systems of interest --bodies are sum of all biomolecules, chemical r(s, & energy transformations. ehavior of all these is governed by thermodynamics. Disease-based alterations in biomolecular str( or organization affect f( and the ability to carry out cellular f( according to the laws of termodynamics. "earn thermodynamic principles to better understand metabolic f(., thereby understanding better the metabolic bases of many disorders. *. Define the term enthalpy, and e(plain what changes in enthalpy imply about a biological system. --enthalpy . f(. related to heat transfer & wor1 e(penditure in the system. . refers to the in enthalpy that occurs during a r(. or in state. For a system eAuilibrium? state 1 state * . = .* .1 'reports heat flow in M0mol+ --negative . = heat lost by system 'favorable+ --positive . = heat gained by system 'unfavorable+ 4. Define the term entropy, and e(plain what changes in entropy imply about a biological system. 'entropy = 2+ --%ntropy measure of disorder. )rdered state entropy, disordered - entropy. --8ncrease in entropy always favored.

--2 reports change in order? state 1state * 2 = 2 * 21 --negative 2 = system becoming more ordered 'unfavorable+ --positive 2 = system becoming less ordered 'favorable+ 5. Define the term free energy, and e(plain what changes in free energy imply about a biological system. --free energy, 9 = hypothetical Auantity how much energy is available for wor1 and allows one to asses reaction spontaneity 'probability+ -- in free energy is favorable. --neg 9 = system in free energy = favorable r( '-., large B2, or both+ --pos 9 = system in free energy = unfavorable r( 'B., small 2, or both+ 7. %(plain the relationship between the N9 value for a reaction and the direction a reaction will move 'slide 11+. -- ! B $ B D & 9 = . -2 so 9 = . - -2 --9 = C, r( at eAuilibrium 'no net change in I!O, I O, I$O, or IDO+ --9 P C, r(. proceeds as written to favor $ and D formation 'spontaneous+ --9 Q C, r(. proceeds opposite to that written to favor ! & form. 'non-spont.+ :. %(plain the relationship between N9 and N9CH. %(plain why the Auantity N9oH is necessary for biological reactions. --9 = 9H B 3- ln DeA 'DeA = I$OIDO 0 I!OI O+ --9 is J standard state, not body environ. 9H is under body conditions as derived under e(perimental conditions, necessary b0c biolog. r(s donHt occur at concentrations used to derive 9 or at the standard temp. and pressure. ;. For a reaction at eAuilibrium 'N9 = C+, understand the relationship between N9 oH and the eAuilibrium constant 'DeA+ for the reaction. --at eAuil., 9 = C, so plugging into above eAuation and rearranging, --9H = -3- ln DeA, if DeA P 1, ln DeA in neg. and 9H is pos. 'more ! & , les $ and D = r( more to left = favoring reactants = B9H+ and ifDeA Q 1, ln DeA in pos. and 9H is neg. 'more $ & D = r(. more to right = favoring products = -9H >. Define the term high energy biomolecule and e(plain their role in driving energetically unfavorable reactions. --serve as energy shuttles, they move energy around to r( sites where it is needed 'do energy transfer+ and drive these energetically unfavorable r(s w0 energy liberated from high energy biomolecule brea1down. @. "ist the two classes of biomolecules that serve as energy transfer molecules. --3educed coenzymes '#!D., F!D.* I#!D/., F!D/.O+ --.igh-energy phosphate compounds 'free energy of hydrolysis Q than -*7 1M0mol+ 1C. Describe the molecules in -able 4.4 on slides *C-** and the relative order of the various molecules in the -able. Rou do not need to 1now the structures. --/38#- -.%2% 2"8D%2? free energy of hydrolysis of phosphate compounds --/%/Q1,4- /9Q!-/QLD/-glucoseQaceteyl $o!Qglucose :-/

11.

%(plain why !-/ is considered the energy currency of the cell, and e(plain why !-/ is such a stable molecule despite having a large energy capacity. Ltilize the e(ample in slide *; as a basis of the e(planation. --energy currency of cell b0c of huge amount of free energy ' 9+ available 'in the 4 phosphate bonds+. L- stable b0c energy of activation is so high, r( is impossible w0o an ez catalyst. !-/-utilizing ezs can do this very rapidly. 1*. Describe the high energy phosphate bonds in !-/, and e(plain why !-/ can participate in multiple reactions utilizing high energy phosphate bonds. --phosphoric anhydride lin1ages have high energyS..T'read about+ --the phosphoric anhydride bonds of !-/ are instable due to neg. chrg on ea. repelling the others. -hus, cleavage of the bond releases energy. !-/ bond cleavage has energy-release than !D/ bond cleavage, but both will release energy. 6ult. r(s utilize !-/, transferring phosphate group to intermediate or protein that is part of energy-reAu. process, thus harnessing the energy released. 14. $ompare and contrast phosphoric anhydride lin1ages and phosphoric-carbo(ylic anhydride lin1ages. --phosphoric-carbo(ylic anhydrides are very energy-rich, more so than phosphoric anhydride lin1ages, b0c of bond strain, electrostatics, resonance. 15. %(plain the principle of coupled energy transfer using !-/ and LD/ to form L-/, L-/ and glucose-1-phosphate to form LD/-glucose, and LD/-glucose to add glucose-1-phosphate units to glycogen. --L-/ energy, glucose 1-/ energy can react together, creating phosphoanhydride bond, giving glucose 1-/ energy. 6a1es pyrophosphate in process which is hydrolyzed, preventing bac1-r( and releasing more energy. 17. Define the term electron shuttle and describe how #!D. and F!D. * 0 F6#.* serve as electron shuttles. Do not be too detailed about electron transport at this point. -his is covered in a subseAuent lecture --electron shuttles carry electrons from one part of metabolism to another. 9oal of metabolism is to move electrons, but they have to be carried. F!DB and #!DB pic1 up .Bs 'e-s+ and carry them as #!D. and F!D.*. 1:. %(plain the role of electron shuttles for energy transfer in mitochondria. 8nclude source of electrons '-$!+ given to the shuttles and the recipients of the electrons '%-$+carried by the shuttles. --e- are transferred from fuel as is goes through -$! to #!DB 'F!DB+ prod. #!D. 'F!D.*+ which shuttles e-s to inner mitochondrial membrane where e- are given to %-$, setting up proton gradient for !-/ synthesis. 1;. "ist the redo( cofactors. --#!DB0#!D., #!D/B0#!D/. --F!D0F!D.* and F6#0F6#.* --coenzyme U --lipoic acid --vitamin $ --tetrahydrobiopterin 18. Describe the redo( reaction !red B o( !o( B red. 2tate what is reduced and what is o(idized initially, and what is reduced and what is o(idized after the reaction. %(plain what happens to ! and to ma1e this change ta1e place. 6

--electron donor 'reducing agent + transfers electrons to electron acceptor 'o(idizing agent+ = reducing agent is o(idized and o(idizing agent is reduced. ! is initially reduced and becomes o(idized when it gives an electron to . is initially o(idized and becomes reduced when it accepts an electron from !. '! Ired.O becomes o(id. by losing e-, Io(id.O becomes red. by pic1ing up e-. -hese two half reactions constitute a conGugate redo( pair. 1@. Describe the redo( potential, and define the standard reduction potential, N% oH. 8nclude the fact that electrons flow spontaneously from a molecule with a more negative to a molecule with a more positive %oH. --redo( potential is the tendency of electrons to be transferred from one biological molecule to another. 6easured in half reactions. <alues are additive, so %H for any conGugate pair is the difference in the % H values of the * half reactions. %H = %H acceptor - %H donor --e- flow spontaneously from a molecule w0 a more neg. to a molecule w0 a more positive %H 'want to move from more '-+ to more 'B++. Lectures "& and '(% )he )*A *ycle 3eference? 2mith, 6ar1s and "ieberman $hapter *C. 1. Describe acetyl $o!. --metabolic pathways lead to activated *-$ acetyl portion of acetyl-$o!. *. Describe the function and the goal of the -$! cycle. --anaerobic metab. resp. for o(idation of acetyl $o! to $)* and .*). --high energy compounds '#!D., F!D.*+ produced during the cycle that lead to the production of !-/. --goal to conserve energy of acetyl $o! o(idation as e- transferred from $ compounds to form #!D. and F!D.* 'reduced form of #!DB and F!D+ --@CV of energy produced by -$! is conserved. 3educed coenzymes are reo(idized by the %-$ in mitochondria w0 prod. of !-/ by o(idative phosph. 4. %(plain the outcome of the -$! cycle in terms of molecules of !-/ and 9-/ produced. 8nclude an accounting of !-/ formed per #!D. and F!D. *. --* $ enter cycle 'acetate group 'acetyl $o!++, * $ released '* $)*+. #o net synthesis 'no in total $ content+. #et yield of energy-containing comp. for ea. cycle is 4 #!D., 1 F!D.*, 19-/. During o(id. phosph. = *.7 !-/ produced by o(id. of ea. #!D., 1.7 !-/ prod. by o(id. of ea. F!D.*, and 1 9-/ prod. = @ !-/ B 1 9-/ = 1C high energy phosphate molecules 5. "ist the > steps of the -$! cycle and state what is the substrate's+, product's+ and enzymes involved in each step. +O)E% you do not need to 1now the chemical structures of every intermediate only those specified on the lecture slides that are discussed in detail by structure and function. --3(s? $ondensations, isomerizations, o(idative decarbo(ylations, dehydrogenations, hydrations via specific enzymes and cofactors. $oupled r(s. --2tep 1? $ondensation of 5-$ dicarbo(ylic acid o(aloacetate ')!!+ w0 acetyl grp. of acetyl $o!. Forms :-$ tricarbo(ylic acid citrate. --2tep *? 8somerization of citrate to isocitrate by aconitase. 7

--2tep 4? Dehydrogenation of isocitrate 1etoglutarate 'hydride removed from the $ bearing the ). & is to #!DB to form #!D.. /roton released. carbo(yl grp on carbon 4 is lost as $)* & db forms b0t $5 and its bound o(ygen. --2tep 5? $)* removed from the carbon, * e- of bond transf. to #!DB #!D.. energy gond b0t coenzyme ! and -carbon is formed. -he dehydrogenation of 1etoglutarate by 1etoglutarate dehydrogenase is $)6/"8$!-%D w0 4 ezs and mult. cofactors. -- 2tep 7? energy of succinyl $o! thioester bond used to generate 9-/ from 9D/ = substrate level phosphorylation for formation of energy comp. w0o use of )*. --ne(t 4 steps convert succinate )!! '* o(idations & prod. of F!D.* & #!D.+ --2tep :? succinate o(id. to fumarate. 1 e- & 1 proton removed from ea. methylene group & transf. to F!DB to form F!D.*. --2tep ;? water adds across db of fumarate, forming malate. --2tep >? )!! regenerated by dehydrogenation of malate. .ydride is transferred to #!DB to form #!D.. 7. Define the term coupled reactions and e(plain why it is important to have tightly coupled reactions in the -$! cycle. -:. %(plain the source of the two carbons that enter the -$! cycle at step 1 'acetate from acetyl $o!+, resulting in the conversion of a 5-$ molecule 'o(aloacetate+ to a :-$ molecule 'citrate+. ;. %(plain how two carbons e(it as $)* during the -$! cycle, resulting in no net increase in carbon content during the cycle. >. %(plain, but do not memorize, the relationships between the various cofactors utilized in the conversion of W-1etoglutarate to succinyl-$o! 'slide 1;+. For e(ample, given the diagram in slide 1;, e(plain that lipoic acid serves as a acyl carrier, transferring the acyl group from -// to coenzyme !, forming succinyl $o!. @. Define substrate-level phosphorylation and use the formation of 9-/ during the reaction of succinyl $o! to succinate as an e(ample. --substrate-level phosph. = formation of high energy phosphate compound w0o the use of )*. -1C. "ist the cofactors that are essential for -$! function. 11. %(plain why overall the -$! cycle is e(othermic even though some reactions are strongly endothermic. --cycle @CV efficient in conserving energy of acetyl $o!. )verall e(othermic b0c three r(s. of -$! have lrg neg. 9oH values 'form. of citrate, D9, succ. $o!+ 1*. %(plain the importance of the two reactions that are largely endothermic 'formation of isocitrate and the formation of o(aloacetate+ in terms of the pool size of citrate 'fatty acid formation+ and malate 'shuttle+. --citrate tells cell whether energy rich or energy poor. --malate build-up tells cycle time to proceed. --citrate and malate 1ey crossover metabolits. 6alate for gluconeogenesis pathway, citrate for fatty acid synthesis pathway.

14. "ist the five molecules of the -$! cycle that lead to other pathways. --citrate to fatty acid synthesis --amino 1etoglutarate to amino acid synthesis or neurotransmitter 'brain+ --succinyl $o! to heme synthesis --malate to gluconeogenesis --o(aloacetate to amino acid synthesis XX ased on cell needs, intermed. can be siphoned off -$! to feed other pathways. 15. Define the term anaplerotic reaction in terms of the -$! cycle and e(plain why these reactions are vital for the -$! cycle to function. "ist the three anaplerotic reactions discussed in class in terms of substrate, -$! intermediate produced and the enzyme used 'e(cept for propionyl $o! to succinate, which uses multiple enzymes+. ---$! intermediates depleted by other pathways. !naplerotic r(s 'filling r(s+ provide -$! intermediates for the cycle. Dey = !"!#$% --pyruvate B $)* )!! by pyruvate carbo(ylase '6)2- 86/.YY+ -17. "ist the two 1ey signals that regulate the -$! cycle. -- '-+ #!D. and 'B+ pyruvate levels = active /D., leads to -$! cycle -- '-+ !D/ and 'B+ $a*B levels = inactive /D. -- * 1ey signals = !-/0!D/ ratio 'energy charge+, #!D.0#!DB ratio 'redo( level+ 1:. "ist and e(plain the 1ey points for regulating metabolic pathways, using the -$! cycle as an e(ample where appropriate. --1;. %(plain why it is crucial to regulate the flu( of pyruvate to acetyl $o!, and why it is eAually important to regulate entry of acetyl $o! into the -$! cycle. 1>. Describe the regulation of pyruvate dehydrogenase in terms of why pyruvate, #!DB and coenzyme ! stimulate the enzyme whereas #!D. and acetyl $o! inhibit the enzyme. !lso, e(plain the effect of reversible pyruvate dehydrogenase phosphorylation by a 1inase and phosphatase and why high !D/ decreases the 1inase activity and high $a*B stimulates the phosphatase. +O)E% the regulation of the 1inase 0 phosphatase pair will be covered in detail in other lectures. 1@. Describe and e(plain the regulation of isocitrate dehydrogenase. 8nclude the modulators and their effect on enzyme velocity. %(plain why having this enzyme as a rate-limiting step is 1ey for -$! function. *C. %(plain the relationship between #!D. levels and the conversion of malate to o(aloacetate. 8nclude a consideration of why it is advantageous to have high malate levels if #!D. is high 'malate shuttle+. --6alate )!! has pos. 9H. #!D. strong signal that stops step. --#!D. = malate needed for gluconeogenesis rather than for -$!. Lecture% Oxidative ,hos-horylation and the Electron )rans-ort *hain

1. -*.

Describe the components of the electron transport chain '%-$+ in terms of integral versus peripherial proteins, lipid-bound molecules versus small metabolites, etc.

Describe how electrons from cytoplasmic #!D. outside the mitochondria are transported into mitochondria by the malate-aspartate shuttle and the glycerol phosphate shuttle. 8n particular, e(plain why the malate aspartate shuttle allows for the production of *.7 !-/s, whereas the glycerol phosphate shuttle results in the formation of 1.7 !-/s. 4. Describe how the #!D. transfers a hydride ion #!D. dehydrogenase to start the electron transport chain reaction. --results in conform. change, acts as proton pump 5. Describe how F!D.* receives electrons, and then transfers the electrons to electron transport chain. 7. %(plain why, as electrons move down the %-$, the carriers become reduced as they o(idize the previous carrier. :. Describe how the energy needs are met to pump protons across the inner mitochondrial membrane from the matri( to cytosolic side. ;. Describe how !-/ is made by the F1FC !-/ase '!-/ synthase+. --slide *;B >. %(plain the e(change of !-/0!D/ at the inner mitochondrial membrane. @. Describe the chemiosmotic theory for o(idative phosphorylation, including how %-$ and !-/ production 'o(idative phosphorylation+ occur simultaneously and are tightly coupled --slide 11 1C. Describe the theory that predicts generation of *.7 !-/s from a molecule of #!D. and 1.7 !-/s from the o(idation a F!D. *, in terms of /0) ratio. --slide 4C, not on e(am 11. %(plain how many protons are needed to ma1e an !-/ '4 protons+ and transfer an !-/ out of mitochondria '1 proton+. 1*. #ame the inhibitors of the %-$ and o(idative phosphorylation. %(plain which %-$ components will be reduced and which will remain o(idized following treatment with the various inhibitors. --slide 44B --inhibitors 1ill target protein, stop f(. --depending on which inhibitor is used, what is reduced, what is o(idized, can !-/ still be producedT --3ontenone, drug, inhibits comple( 8, so when #!D. dumps its hydride and becomes reduces. ut electrons canHt be passed on, so comple( 888 and on remain o(idized b0c no more e- coming through. $omple( 88 is still active, so e- still coming in through there, so %-$ can still wor1 and !-/ can still be made. 9radient ends up w0 : .B to intermembrane space, not 1C, but still have gradient --6alonate, drug, inhibits comple( 88, so e- entry bloc1ed at comple( 88, but comple( 8 can still bring in e-s, 1eeping %-$ going and !-/ produced. 9radient ends up w0 : .B to intermembrane space, not 1C, but still have gradient

10

--!ntimycin-! inhibits comple( 888. #!D. comes in and gives e- to $oU, as does comple( 88. $oU wants to pass them on, becoming more reduced, bac1s up, but comple( 8 now completely reduced and comple( 88 now completely reduced b0c canHt pass e- on. #!D. and fumarate can no longer pass on e-. 2o, end up with collapsing of proton gradient and canHt ma1e !-/ b0c stops %-$. %verything before comple( 888 becomes completely reduced and everything after is eventually o(idized. --carbon mono(ide, azide, cynide 1ills off comple( 8<, same scenario as w0 comple( 888, even more severeT $) binds w0 such affinity, almost impossible to replace it. --)ligomycin inhibits !-/ synthase 14. Describe the compounds 'i.e., uncouplers+ that cause lea1age of protons through the inner mitochondrial membrane and thus prevent formation of a proton. --!-/ synthesis reAuires intact inner mitoch. membr. w0 .B gradient --any agent that allows .B to lea1 bac1 into matri( uncouples the %-$ 15. %(plain why electron transport occurs in the presence of an uncoupler, but not !-/ production. 17. %(plain how the rate of o(idative phosphorylation is controlled by the cell. 1:. Describe cyanide poisoning. '.int? $yanide binds to Fe 4B in $ytochrome aa4 resulting inhibition of respiration and stoppage of energy production, leading to rapid death+. 1;. Describe malignant hyperthermia. '.int? -he maGor inhalation anesthetics halothane, ether, and metho(yflurane trigger a reaction in susceptible people, which results in the uncoupling of o(idative phosphorylation from electron transport. !-/ production decreases, heat is generated and temperature rises mar1edly, e(cessive $)* production leads to respiratory acidosis. 18. Describe how acute myocardial infraction can destabilize the cellular membrane. '.int? coronary occlusions occur lac1 of o(ygen lac1s of !-/ for contraction and maintenance of membrane integrity.+ Lecture '"% Glycolysis Generates A), from Glucose 3eference? 2mith, 6ar1s and "ieberman $hapter *1. 1. /rovide a general overview of glycolysis in terms of the goal of the pathway and the outcome in !-/ and #!D.. --glucose is universal fuel for human cells. 9oal of cell is to generate !-/ and reducing eAuivalents from glucose. glycolysis generates * moles #!D. and * moles !-/ per mole of glucose. so glycolysis is the pathway for o(idation of glucose to pyruvate. #et !-/ produced independent of -$! cycle 'anaerobic !-/ production+, then 5 !-/ produced and * #!D. produced. #!D. produced 'reduced cofactor+ leads to !-/ production in mito. Ehen pyruvate enters -$! cycle 'aerobic+lots of !-/ '4:-4> moles !-/ per mole glucose+. *. "ist the four fates 'i.e., products+ that can be formed from pyruvate. --alanine, oaloacetate, acetyl $o!, lactate 4. "ist the two cell types that are dependent on glucose for energy. --3 $s no mitochondria

11

--brain cells lots of )* and mitochondria, but cells lac1 machinery to use fatty acids for fuel. plus, fatty acids donHt cross blood-brain barrier 5. "ist the glycolysis intermediates that are used by other pathway. --pentose phosphate pathway uses ribose 7-phosphate = sugar for nucleotides -7. /rovide a summary of the glycolytic pathway 'slide ;+. --occurs in cytosol, cleaves one mole of glucose to * moles of pyruvate. --glucose B * #!DB B */i B *!D/ * pyruvate B *#!D. B 5.B B *!-/ B *.*C --overall net neg. 9H, which drives pathway forward --reversal of glycolysis is gluconeogenesis, reAu. energy since net 'B+ 9H :. "ist and describe the two phases of glycolysis. -;. %(plain the concept of a committed step in a pathway, using the conversion of glucose to glucose-:-phosphate as an e(ample. --use of 1 !-/ to phosphorylate glucose. traps glucose inside cell 'commits glucose to cell+ >. %(plain the importance of glucose-:-phosphate as an intermediate in glycolysis and other metabolic pathways. -- if energy needed, go forward into glycolysis, if have plenty, could go to glycogen synth., could go to pentose phosphate pathway or other pathways, too. @. $ontrast and compare the function of he(o1inase and gluco1inase, in terms of the tissues where these two enzymes are found. -1C. %(plain why the action of phosphofructose-1 '/FD-1+ is considered another committed step in glycolysis, and why this enzyme specifically commits glucose to glycolysis. 11. %(plain why it ma1es sense that /FD-1 is a rate-limiting enzyme for glycolysis, and why this enzyme is highly regulated. 1*. Describe how four molecules of !-/ are produced by substrate-level phosphorylation at two steps during glycolysis, and name the high energy phosphate compounds used to drive !-/ formation. 14. Describe the reaction that results in the production of #!D. in glycolysis. !lso, e(plain why #!DB is reAuired for glycolysis to occur. 15. %(plain the function of the malate0 aspartate and glycerol-4-phosphate shuttles for e(changing #!D. 0 #!DB reducing eAuivalents across the inner mitochondrial membrane. 17. %(plain the glycerol-4-phosphate and malate0 aspartate shuttle systems, including the moles of !-/ that are produced per #!D. shuttled in each system. 1:. %(plain the conditions that favor 'or reAuire+ conversion of pyruvate to lactate, and how this is an important source of #!DB for additional glycolysis 'i.e., #!DB is generated, which is needed for glycolysis+. 1;. "ist the fates of lactate in cells that can utilize it. e sure to understand the method by which lactate is used depends on the metabolic state of the cell. For e(ample? if glucose is needed, lactate is used to ma1e glucose. 8f !-/ is needed, lactate is used to generate !-/, etc. 1>. %(plain the $ori $ycle and why this is crucial for the function of red blood cells.

12

Lecture ''% A), from .atty Acids and /etone Bodies 3eference? 2mith, 6ar1s and "ieberman $hapter *4 1. Define Z-o(idation of fatty acids and describe in general the purpose of this pathway. *. Define 1etone bodies and understand what tissues ma1es them, and what tissues can and can not utilize themSand why. 4. "ist the four types of fatty acid classification by chain length. 5. %(plain fatty acid activation and why this is a necessary step for Z-o(idation. "ist the sites in the cell where the various forms of fatty acids are activated. 7. %(plain the mechanism of long-chain fatty acid transport from the cytosol to the mitochondrial matri(. %(plain this at the level of detail provided on slide @. :. %(plain why newly synthesized fatty acids are not immediately transported into the mitochondrial matri(. ;. Describe the step by step process and outcomes of fatty acid o(idation, using palmitoyl $o! as an e(ample. 8nclude the energy yield of !-/. +O)E% 6a1ing the math wor1 out for moles !-/ produced per *-carbon unit is tric1y at best. Must state that more than 1CC !-/ are produced for palmitoyl-$o! compared to about 4> for glucose. >. %(plain that propionyl $o-! is the final carbon product of odd-chain fatty acid o(idation, and that this intermediate can produce glucose. %(plain that propionyl $o-! can lead to succinyl-$o!, which can enter the -$! cycle and become o(aloacetate, then on to glucose formation via gluconeogenesis. ---o(idation occurs Gust as w0 even chain until 7-$ left. -hen produces 1 more acetyl $o! and one propionyl $o! '4 carbon fatty acyl $o!+. /ropionyl $o! is carbo(ylated to methylmalonyl $o!, which is converted to succinyl $o! '-$! intermediate+. 2uccinyl $o! can lead to glucose formation via gluconeogenesis. @. $ompare and contrast fatty o(idation for saturated versus unsaturated fatty acids. %(plain how the fatty acid double bond is reconfigured 'if necessary+ to allow o(idation to proceed. 8n particular, understand how and why the presence of double bonds reduces the yield of !-/. ---Fatty acid db must be in proper config. b0t and carbons. 8f not, e(tra ezs rearrange db, which reAuires energy, yield of !-/. 1C. Describe the o(idation of very long fatty acids in pero(isomes. -11. "ist the three 1etone bodies produced by the liver and which is most prevelent, and e(plain which tissues use these 1etone bodies and under what conditions. --acetoacetate, -hydro(ybutyrate, acetone 'synthesis prim. in liver+ --used by muscle 'and brain when levels are high+, 1idneys, D s used in long fasts and starvation states ---hydro(ybutyrate most prevalent

13

1*.

"ist the metabolic conditions that stimulate formation of 1etone bodies. 8n particular, e(plain why high levels of !cetyl $o! signals the formation of 1etone bodies. --stim. when blood levels of F!s. -14. %(plain the time course for the increase of 1etone bodies in the blood during fasting 'slide *1+. -15. %(plain the synthesis of acetoacetate from acetyl $o!, and the production of DZ-hydro(ybutyrate and acetone from acetoacetate, and also the reverse reactions bac1 to acetyl $o!. 17. %(plain why more !-/ is produced from the o(idation of D-Z-hydro(ybutyrate than the o(idation of acetoacetate. 1:. %(plain why converting the energy of fatty acid o(idation to 1etone bodies is advantageous compared to simply o(idizing the fatty acids all the way. 1;. %(plain the metabolic conditions that regulate fatty acid o(idation and 1etone body production. Lecture '0 and '1% Oxygen Metabolism and Oxygen )oxicity 2 Misra 3eference? 2mith, 6ar1s and "ieberman $hapter *5 Free radicals and reactive o(ygen0nitrogen species are involved in a variety of biological phenomena such as aging, .8<0!8D2, malaria, diabetes, cancer, malnutrition, cardiovascular dysfunctions, and responses to agricultural e(posure to pesticides, radiation inGury, ischemia-reperfusion inGury, and neurodegenerative diseases. !t the end of these two lectures students should be able to answer the following Auestions? 1. Describe the electronic structure of o(ygen and spin restriction. Ehy do humans not spontaneously ignite in the o(ygen rich atmosphereT

---o(ygen spin restriction *. Define free radicals and describe the step wise reduction of o(ygen to form * molecules of water and the reactive intermediate products. --free radicals are molecules w0 unpaired e-, very reactive, short half-life, can initiate chain reactions. --)* gains an e- and becomes supero(ide ') 2-), gains another e- and 2 H+ to become H2 2 h!drogen "ero#ide. $ains a third e-, %orming H2 and a h!dro#!& radica& ' H( ) *er! reacti*e+ most "otent o#idant ,no-n). .he 4th e- is trans%erred in, a&ong -/ a H+, and the 2nd H20 is %ormed. 4. Describe o(idative stress. Ehat are various stimuli that increase the generation of 3eactive )(ygen 2pecies '3)2+T slide 18? --normal metabolism --inflammation --radiation

14

--aging --high p)* --2mog ')4, #)*+ --$hemicals and drugs --reperfusion inGury 'following ischemic bloc1+YYY can = death in spite of successful surgery to remove occlusion all produce reactive o(ygen species )*-, .*)*, ).X 5. Describe the biological sources of 3)2 and the cellular damage that occurs by 3)2 attac1. see slide 22? slide 17? --endoplasmic drug metabolism, mitochondrial electron transport chain, ionizing radiation, enzymes, phagocytes, auto-o(idation of drugs and biomolucules 'hemoglobin, epinephrine, thiols, phenylhydrazine+ --protein damage --membrane damage --D#! damage --"ipid pero(idation --massive influ( of $a*B --permeability --cell swelling --mitochondrial damage /ar1insonHs disease )* ed to pero(ide by 6!), going to ).X radical 'by Fenton r(.+ and mitochondria prod. a )*-, Goining w0 #) to form 3#)2. 3esults in lipid pero(idation, protein o(idation, D#! strands brea1 lipofuscin. -hus neuronal degeneration and reduced dopamine release. 7. Describe the reactions that generate the deadly hydro(yl radicals. '.int? the Fenton 3eaction and the .aber-Eeiss 3eaction+ :. Describe the lipid pero(idation chain reaction - the initiation, propagation, degradation and termination steps. --lipid pero(idation rancid. --initiation long chain carbonhydrate '".+ B ).X "X B ).. -hus ).X 'hydro(yl radical+ is initiator, producing hydrogen abstraction --propagation continues chain reaction until all ". is o(idized. "X B )* "))X and "))X B ". ")). B "X --degradation --termination "))X B "X ")). B ". = no more radical = termination or "X B <it% ". B <it%X and <it%X B "X ". B <it%o( = termination antio(idants help in process. --6alondialdehyde shows up in degradation stage and can be detected in blood or urine 'via color change when added to thiobarbituric acid+, indicating free radical tissue damage = o(idative stress w0in body. ;. Ehich amino acids in proteins are particularly susceptible to 3)2 attac1T --/ro, .is, !rg, $ys, 6et more prone to o(idative stress >. Define respiratory burst during phagocytosis. -@. Describe cellular defenses against o(ygen to(icity. 15

--supero(ide dismutase, catalase, 2)D, glutathione pero(idatse, 92., vitamin $, %, carotene 'in !+ are antio(idants, hemosiderin ferritin 'removes free iron, protecting against free radical o(idation+, antio(idant enzymes 'supero(ide dismutase '4 forms cytosolic, mitochondrial, e(tracellular+, catalase, glutathione pero(idase, glutathione reductase+ --see slide 51 1C. Ehy does the cell need such a high content of 6n-containing 2)DT 11. Describe which organs contain the highest activities of the antio(idant enzymes. '.int? where mitochondrial and pero(isomal contents are high and $ytochrome /57C enzymes are found in abundance+. -1*. -he levels of o(idative damage to mitochondrial D#! are at least 1C-fold higher than nuclear D#!, and increase with a personHs age. EhyT --nuclear D#! protected by histones --mtD#! located near inner membrane of mitochondria where %-$ is located and 3)2 generated from %-$ 'at $oU+ doesnHt have far to go to attac1 mtD#! -- w0 age b0c SS 14. Describe the electron lea1s in the mitochondrial electron transfer chain. 15. Ehat is nitric o(ideT #itric o(ide is both essential to life and to(ic. %(plain. 17. Describe the reaction catalyzed by nitric o(ide synthase. Ehat are different isoforms of nitric o(ide synthaseT Ehat are their specific roles in the bodyT --4 isoforms inducible, neuronal, endothelial 'inducible+ --nitric o(ide synthase #) )#)) pero(ynitrite --urea cycle, arginine citrulline B #) --e#)2 and n#)2 regulated by $a*B act as neurotransmitter and hormone --i#)2 present in many cells 'induction of gene+ responsible for bul1 of 3#)2 production phagocytosis, inflammation, D!# dmg, protein and lipid damage. 1:. Describe the amino acid composition of glutathione '92.+ present in all mammalian cells 'many non-mammalian cells too+ e(cept the neurons. 1;. Describe the conseAuences of 92. deficiency in new born and in adults. --death in newborns, cataract and mitochondrial swelling in adults. 1>. Describe in detail the $ytochrome /57C system, which hydro(ylates many physiological compounds, including steroids, fatty acids and many (enobiotics 'drugs, carcinogens and environmental agents+. 1@. -he drug metabolizing enzyme utilizes #!D/., rather than #!D. as a source of electron. Ehy this choiceT *C. Ehat would be benefit of administrating therapeutic doses of antio(idants to patients immediately after a car accident, before cardiac bypass surgery and before administrating do(orubicin '!driamycin+T *1. Ehy there is an e(cess production of 3)2 during ischemia0reperfusion inGury in heart and in intestine. Describe the mechanism of action of nitroglycerin. **. /ar1insonism pathogenesis may be multi-factorial. 8n the early stage, a monoamine o(idase beta-inhibitor is used. Describe its mechanism of action. Lecture '#% 3ormonal Regulation of Metabolism 16

3eference? 2mith, 6ar1s and "ieberman $hapter *: 1. -*. %(plain the concept of balance between tissue needs for fuel and fuel availability.

%(plain that hormones respond to imbalances and stimulate or inhibit systems to restore balance. --hormones carry messages to individual tissues about the state of the body and supply of fuel or demand of nutrients, 4. %(plain the concept of counter-regulatory hormones, and e(plain this concept using the fundamental effects of insulin and glucagon as an e(ample. --effect of ea. hormone counteracts the effects of the other hormone. 8nsulin and glucagon are opposites. released in response to circulating levels of fuel in blood. insulin is the boss hormones, glucagon responds 'when insulin comes out to play, glucagon Gust goes away+ --8nsulin anabolic, signals fed state, produced by pancreatic -cells, promotes storage of fuels or util. of fuels for growth. 3elease of insulin dictated by blood glucose level. --9lycogen catabolic, signals fasted state, 1 insulin counter-regulatory hormone. produced by cells. promotes fuel utilization, acts on liver, adipose tissue, no effect on muscle, release depends on ! 2%#$% of glucose, insulin 'glycogen level lowest after a meal+. 9lucagon !2 insulin 'highest during prolonged fast & in starved state+. 5. Describe changes in glucose levels, insulin levels, glucagon levels and nitrogen levels in the blood over 4-5 hours following a high carbohydrate versus a high protein meal. --slide 1* snuggly slide print out 7. %(plain the relationship between endocrinology and metabolism, and that signals are communicated between various organs to utilize and store e(cess fuel or to mobilize fuel reserves when fuel is low. -slide *1 :. %(plain that the relative levels of insulin versus glucagon following a meal determine the flu( through the various metabolic pathways. 8nsulin turns off glucagon-stimulated pathways and glucagon turns off insulin-stimulated pathways. ;. %(plain metabolic pathways stimulated by glucagon signaling 'slides 14 and 17+. 8n particular, e(plain the balance between glycogen synthesis and glycogen brea1down, and how glucagon stimulates one aspect 'brea1down+ whereas the absence of glucagon 'i.e., the presence of insulin+ stimulates the other aspect 'glycogen synthesis+. >. %(plain signal transduction by insulin, and its action to counteract the effects of glucagon on the cell 'i.e., slide 14 and 1@, and undoing the glucagon effects in slide 17+. +O)E% 8t is not necessary to understand insulin action at the level of slide 1:. -his is Gust to help review the details of insulin action. @. %(plain the fight or flight response to epinephrine that elicits metabolic changes in glycogen brea1down in muscle versus liver. 8n muscle, this is the same response pathway as the glucagon pathways. 8n liver, epinephrine wor1s 17

1C.

through the phospholipase $ pathway. %(plain this at the level of detail presented in slide *5. %(plain that the regulation of glycogen brea1down during e(ercise is by a similar mechanism to the fight or flight pathways, but the signal initiating the pathway during e(ercise is different 'i.e., $a*B levels rise in response to nerve impulses during e(ercise+.

Lecture '$% 4igestion and Absor-tion of *arbohydrates 3eference? 2mith, 6ar1s and "ieberman $hapter *: 1. "ist the carbohydrates found in food. --oligosaccharides polymer containing *-1C monosaccharides --dietary disaccharides are most common --prebiotic carbs. oligosaccharides that selectively support the growth and activity of good intestinal microflora 'galacto- & fructo- oligo., soy oligo., inulin. *. "ist the common dietary carbohydrates. --plant starches, sucrose from fruits, vegs., glucose, fructose from fruits, honey, lactose from mil1 products, dietary fiber, high fructose corn syrup. 4. %(plain the difference between amylose and amylopectin, since both are derived from starch. --both products of starch, both digested by -amyhlase, maltase, isomaltase. --diff? amylase disacc., amylopectin double disacc. 5. Describe the function of glycosidases. --ezs that brea1 down carbs. to simpler sugars 'saccharidases, lactase, etc+ 7. #ame the specific enzyme that brea1s down internal W-1,5 glycosidic bonds in a starch. #ame the various products produced by this enzyme, and list which starch bonds are not affected by this enzyme. :. %(plain why humans cannot digest cellulose. ;. "ist the saccharidases that degrade each common dietary sugar. >. Define the maGor mechanisms of sugar absorption by intestinal epithelium. @. $ompare and contrast glucose transport for neural versus non-neural tissues. 8n particular, e(plain why this difference ma1es glucose upta1e through the bloodbrain barrier much slower. --neural endothelial cells tightly sealed, creating barrier. !nything that goes thru has to go thru transporters. 9lucose transporters, but no fat transporters. --no 1C. %(plain lactose intolerance. --lactase deficient = no brea1down of lactose = lactose in lumen = becomes bacterial food = methane, lactic acid production = gas, diarrhea. 11. "ist and describe the three classes of starch. --rapidly digestible starch --slowly digestible starch --resistant starch so tightly pac1ed that body canHt brea1 down --slow glucose prod. from 2D2 and 32, blood glucose levels donHt rise as Auic1ly from these foddstuffs. -hus, some starch foods actually have glycem. inde(

18

--3esistant starch implicated as food source for bacteria in lower intest. tract. 1*. Define the term dietary fiber and describe the two types of dietary fiber. --fiber = sum of polysacc. and lignins not digested by human 98 tract --soluble vs. insoluble 14. %(plain probiotics what they are, what they do and why they are good for colon health. 8nclude a consideration of probiotic versus pathogenic bacteria. --slide *C, *1, ** 15. Describe inulin and the three types of fructooligosaccharides produced when bifidobacteria brea1 down inulin. %(plain why these compounds called prebiotics. --slide 1@ Lecture '&% Glycogen Metabolism 3eference? 2mith, 6ar1s and "ieberman $hapter *> 1. %(plain the need for glycogen as a glucose storage mechanism. --glycogen = reservoir of glucosyl units for !-/ generation from glycolysis. --muscle metabolizes glycogen much more rapidly than F!s --F!s not metabolized under anaerobic conditions --!cetyl $o! 'produced by F! brea1down+ canHt be converted to glucose --Free energy of r(. of !cetyl $o! to pyruvate too largely positive *. %(plain that the body stores e(cess glucose as glycogen to maintain a ready supply of glucose, rather than Gust convert all the e(cess glucose to triacylglyerols '-9s+, which canHt be converted bac1 to glucose. --#eed glycogen to provide glucose for 3 $s, brain. 4. Describe the structure of glycogen. %(plain the logic of having a highly branched glycogen structure, and why the one reducing end is bloc1ed. --branched glucose polysaccharide = glucosyl units lin1ed by -1,5 glycosidic bonds w0 -1,: branches every >-1C residues, present in all tissues as molecular weight polymers, collected together in glycogen particles --only one residue has a reducing end, attached to glycogenin protein, other ends of chain are non-reducing ends. -- ranched str(. allows for tight pac1ing of glucose, rapid degradation0synthesis, ez. ability to wor1 on several branches at the same time. 5. 8dentify the W-1,5 and the W-1,: glucose lin1ages in glycogen and e(plain how they contribute to the structure and function of glycogen. -7. %(plain the glycogen is bro1en down to glucose-1-phosphate units, which can be used immediately by tissues 'enters directly into glycolysis+. --glycogen degraded to glucose 1-phosphate glucose :-phosphate used in glycolysis in muscle. :. %(plain why muscle and liver have relatively large amounts of glycogen, but other tissues maintain only a small amount of glycogen. --

19

;.

%(plain each step in the glycogen synthesis and degradation pathways, as outlined on slide >, at the level of detail provided on subseAuent slides 'esp. 1*, 15 and 17+. --glycogen degradation? debrancher ez. brea1s -1,: lin1ages. glycogen phosphorylase attached phosphate glucose-1-/. --primarily in liver. glycogen phosphorylase cleaves single glucose units by transferring / from !-/ to anomeric $ of glucose, brea1ing -1,5 glycosidic lin1age. 3eleases glucose 1-/ 'converted to glucose :-/ by phosphoglucosemutase and enters other pathways+. 8n liver, glucose-:-/ dephos. by glucose-:-phosphatase & transported out of cell by gluco1inase. --glycogen phosphorylase canHt act on glycosidic bonds of 5 glucoses adGacent to brach 'doesnHt fit in active site+, so debrancher ez SS --2ynthesis? 'mostly adding to glucose units to e(isting glycogen molecules+ glucose1-/ B L-/'LD/-glucose pyrophosphorylase+ LD/-9. LD/-9 --anomeric $ of glucose on LD/-9 forms -1,5 lin1age w0 $-5 on glucose at non-reducing end of glycogen chain, displacing LD/ and chain length. Ehen chain K11 residues long, a :-> residue piece is cleaved from end of chain and reattached to a glycosyl unit by an -1,: bond, forming a branch point. %a. branch is lengthened until they are cleaved to form new branches, etc. >. %(plain the regulation of glycogen degradation by glucagon signaling, as shown in slide 1;. -@. %(plain that is it is necessary to activate glucose-1-phosphate to LD/-glucose to ma1e it reactive and able to bind to the glycogen. !lso understand that LD/glucose is a common intermediate that can be used by other pathways. 1C. Describe how most cells retain glucose 1-phosphate for their own use, but liver converts this to free glucose for e(port into the blood 'i.e., produces glucose for the body+. %(plain why cells other than liver cells canHt produce free glucose. 11. %(plain that glycogen brea1down produces mostly glucose-1-phosphate, but that one glucose unit is produced for every branch point that is degraded. 1*. %(plain lysosomal degradation of glycogen to glucose. 14. Describe how high levels of !6/ in muscle lead to an activation of glycogen phosphorylase and increased glycogen brea1down. $ompare this activation to that observed when glycogen phosphorylase is activated by phosphorylation by /D!. Lecture 0(% )he ,entose ,hos-hate ,athway and 5nterconversion of ugars 3eference? 2mith, 6ar1s and "ieberman $hapter *@ 1. Describe the nature of the pentose phosphate pathway and its goals. "ist the final products besides #!D/.. --pent. phosph. pathway alt. pathway for glucose :-phosphate utilization, a shunt. --

20

*. 4.

5.

7. :. ;. >.

@. 1C.

11.

"ist some metabolic pathways depend on the #!D/. produced by the pentose phosphate pathway. %(plain what will happen to these pathways if the pentose phosphate pathway is interrupted 'common test Auestion+. "ist and describe the two phases of the pentose phosphate pathway. %(plain which phase is irreversible 'and why+ and which phase is completely reversible. "i1ewise, e(plain why the second phase of the pentose phosphate pathway can operate even if the first phase is shut down. --o(idative phase --non-o(idative phase 7 rearrangement & transfer r(s that occur in * freely reversible parts part 1 = * isomerizations of ribulose 7-phosphate, part * pentose molecules are converted to intermediates of the glycolytic pathway. 3(s. of phase 1 are irreversible, only occur if #!D/B is available ' #!D/., #!D/ ratio stiulates the pathway, #!D/. inhibits pathway 'shuts down pt 1+ 3(s. of phase * are all reversible can f(. independently to meet metabolic need for ribose 7-/ or for gucose metabolism intermediates. Ehen #!D/. levels are normal, phase 1 is shut down, but step * can operate as needed. %(plain the importance of the first step of the pentose phosphate pathway 'glucose-:-phosphate dehydrogenase+ being the rate-limiting step of the pathway. --o(idative phase of pentose phosphate pathway = maGor source of #!D/. in cells. 9-:-/ dehydrogenase 1ey ez. in #!D/. production. --9-:-/ is o(idized in o(idative phase, reducing #!D/B. TTTT %(plain why the first and third steps of the pentose phosphate pathway are strongly inhibited by #!D/.. "ist some of the 1ey intermediates produced in the second phase of the pentose phosphate pathway that can be used by the cell. %(plain that the flow of the second phase is determined by cellular needs. %(plain that the final products of the pentose phosphate pathway are intermediates of glycolysis that reenter that pathway. %(plain the role of #!D/. in counteracting reactive o(ygen species in red blood cells. %(plain what could happen to these cells if insufficient #!D/. is produced by the pentose phosphate pathway. --#!D/. helps maintain glutathione in an active form. 9lutithione pero(idase SS "ist the three common sugars found in the human diet 'not sucrose, but the constituents of sucrose+. --fructose, galactose, glucose %(plain that any sugar we need for glycoprotein or glycolipid synthesis can be made from glucose, and any sugar we consume can be made into glucose. --all sugars we need for various uses can be synthesized from dietary glucose. --avail. of other sugars in our diet provides 1ey starting material for synthesis of carb. derivative in glycolipids, glycoproteins, proteoglycans '9!9s+, etc. "ist and describe the steps that produce glyceraldehydes-4-phosphate and dihydro(yacetone phosphate from fructose in liver. %(plain the 1ey role !ldolase plays in this process and understand how defects in this enzyme can affect fructose metabolism.

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14. 15.

17. 1:. 1;. 1>.

-%(plain how fructose is utilized by non liver cells. --phosph. at $: by he(o1inase to form fructose :-/, which enters glycolysis. .e(o1inase phosphorylates glucose preferentially, but will wor1 on fructose. --* moles !-/ used per mole fructose to reach glycolytic intermed., sim. to gluc. Describe how fructose can be synthesized from glucose. --fructose prod. from glucose glucose reduced to sorbitol, which reduces the aldehyde group to an alcohol. 2orbitol is o(idized J $-* = fructose. %(plain how galactose is processed and converted to LD/-glucose upon entering the cell. %(plain that LD/-galactose and LD/-glucose are easily interconverted based on cellular needs. -"ist and describe some of the uses for LD/-galactose and LD/-glucose. Define glucuronate and glucuronides and e(plain what their functions are in cells. "ist some of the common fates of LD/-glucuronate in cells. $ompare and contrast classical versus non-classical galactosemia and e(plain why the failure of the two source enzymes causes problems for the cell.

Revised Lecture Ob6ectives Lecture 07% Glucose Metabolism in the Liver 3eference? 2mith, 6ar1s and "ieberman $hapter 41 1. Define gluconeogenesis and e(plain the goal of the pathway. --ma1ing glucose from precursors --supplies glucose to the blood over the long term, as long as precursors avail. *. %(plain the concept of the bypass enzymes and how they are used to avoid energetically unfavorable reactions in gluconeogenesis. --gluconeogenesis is reverse of glycolysis. but 4 metabolically irreversible r(s. of glycolysis have to be bypassed. 4. "ist the three starting materials for gluconeogenesis and e(plain how they are produced by the body. --"actate --9lycerol --!mino acids, particularly alanine 5. "ist the three bypass reactions in gluconeogenesis and e(plain the reason for each step. 7. "ist and describe the steps in the first bypass reaction that ta1es pyruvate to phosphoenoylpyruvate '/%/+. 8nclude an e(planation of why a shuttle system is reAuired to move malate out of the mitochondria in this bypass and why energy is reAuired to form /%/ from pyruvate. :. %(plain that two molecules of pyruvate '4 carbons+ are reAuired to produce on molecule of fructose 1,:-bisphosphate ': carbons+.

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%(plain why it is energetically more favorable to dephosphorylate fructose 1,:bisphosphate and glucose 1-phosphate using phosphatases rather than the reverse reaction of a 1inase in the second and third bypass steps. >. "ist the energy reAuirements for gluconeogenesis and e(plain where the energy eAuivalents come from. @. 3eview levels of glucose, insulin, and glucagon in the blood within several hours after a high carbohydrate versus high protein meal. %(plain the cyclic nature of these levels over the course of *5 hours with three meals at regular intervals. -1C. %(plain how liver, muscle and adipose cells respond to the presence of glucose. $onsider how brain cells respond, too. 11. %(plain the control points in glycolysis 'he(o1inase or gluco1inase, /FD-1+ that affect whether glucose is utilized immediately for the cell 'glycolysis+ versus some other pathway. 1*. Define adenylate charge 'including the normal range for cells+ and e(plain why catabolic 0 anabolic processes are inhibited 0 stimulated by high adenylate charge. 14. %(plain why the !6/ level in a cell is the most potent factor for determining changes in adenylate charge. 15. "ist the allosteric activators of /FD-1 '!6/ and fructose-*,:-bisphosphate+ and e(plain why it ma1es sense for these molecules to stimulate /FD-1. 17. %(plain why !-/ at a low level stimulates /FD-1, but higher levels of !-/ inhibit /FD-1. 1:. %(plain Figure 41.14 showing the sources of glucose during the fed, fasted and starved states during the hours and days following a meal. %(plain, in particular, the overlap between the various plots. For e(ample, note the gluconeogenesis starts and increases rapidly as glycogenolysis decreases, so that total blood glucose remains fairly constant. 1;. %(plain the signals that inhibit glycolysis but stimulate the gluconeogenesis bypass reactions during fasting. %(plain that without these counteracting controls, these reactions would be futile steps that would burn !-/ rapidly. 1>. %(plain the effect of fructose *,:-bisphosphate on /FD-1 and fructose 1,:-bisphosphatase, and the subseAuent effect on the interconversion of fructose-:phosphate to fructose-1,:-bisphosphate. 1@. Describe phosphofructo1inase-* 0 fructose *,:-bisphosphatase 'i.e., the bifunctional enzyme+ as an enzyme that has two functions? it can act as a 1inase that produces fructose *,:-bisphosphate from fructose 1-phosphate or it can act as a phosphatase that produces fructose 1-phospate from *,:-bisphosphate. *C. %(plain the cellular signals that control the function of the bifunctional enzyme. dinsulin is low, which favors the phosphatase function, versus dephosphorylation of the bifunctional enzyme when insulin is high and glucagon is low, which favors the 1inase activity of the enzyme. *1. %(plain why gluco1inase can readily produce glucose :-phosphate when glucose is high, but almost no glucose :-phosphate is formed using glucose produced by gluconeogenesis.

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