Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
Heinz Schwarzkopf
hs0215@live.unthsc.edu
LGT Disclaimer:
LGTs are designed by CAP tutors. They are not meant to replace information you
receive in class or textbooks. While it has been reviewed for typos and errors, you
must always remember that your professor and the material given by them
supersede any information presented in the LGT. The LGT serves as a review, and
are not meant to be an all-inclusive presentation.
Catabolic pathway:
glycolysis
Catabolic
Anabolic
Regulation
Product E
In feedforward stimulation, a
substrate stimulates the pathway by
which it is utilized.
6CO2 + 6H2O
When you see Dehydrogenase that is a clue that you are
doing a redox reaction. Then remember that an
NADH/NADPH/FADH2 is either used or produced.
2.
3.
4.
5.
Electron Carriers
Both coenzymes (NAD+ and NADP+) undergo reversible reduction of
the nicotinamide ring.
What can niacin deficiency lead to?
a) Rickets
b) Diarrhea
c) Sterility
d) Dermal sensitivity
FAD/FADH2
Flavoproteins are enzymes that catalyze oxidation-reduction
reactions using either FMN or FAD as coenzyme. FAD or
FMN are coenzyme derived from the vitamin B2, riboflavin.
Adenine
N-glycosidic bond
In a Healthy Cell
NAD+
NADP+
NADH
NADPH
Metabolism
All major nutrients (carbohydrates, fat, protein, etc.)
are degraded to acetyl coenzyme A (acetyl-CoA)
Think of acetyl-CoA as the center molecule of metabolism
Monosaccharides
From the breakdown of glycogen
Glucose synthesis via amino acids
Glucose synthesis via acetyl CoA
Glucose can be obtained from all of these
sources
Glut1
Glut2
Glut3
Glut4
Glut5
21
23
Brain
Red Blood Cells
Liver
Adipose Tissue
Skeletal Muscle
6 - Carbon sugar
first priming reaction
Step 1
Step 4
3 - Carbon sugar
3 - Carbon sugar
Step 5
2
Step 6
2
Step 7
2
Step 8
2
Step 9
2
2
No oxygen needed
50
Cytosolic NADH+
- It must be re-oxidized:
1. anaerobic conditions: by conversion of pyruvate to lactate by
lactate dehydrogenase, and NADH is re-converted to NAD+,
without oxygen participation.
Regulation of Glycolysis
- Phosphofructokinase-1 (PFK-1), is the most important regulated
enzyme of glycolysis. PFK is:
- stimulated by insulin and inhibited by glucagon (in the liver).
- inhibited by citrate as well as by low pH.
- inhibited by ATP and stimulated by AMP and ADP.
- Hexokinase / Glucokinase: also regulated
- Pyruvate Kinase: also regulated. Inhibited by ATP.
Phosphofructokinase-1(PFK-1)
- Is the most important regulated enzyme of glycolysis.
- Commits glucose to pyruvate in glycolysis.
- Activated by:
- AMP
- Fructose 2,6-bisphosphate synthesized by PFK-2
- levels determined by insulin/glucagon
- levels determined by fructose 6-phosphate in muscle
- Inhibited by:
- ATP
- Citrate
- Glucagon
Pyruvate Kinase
In glycolysis, step 10, phosphoenolpyruvate is converted to
pyruvate by pyruvate kinase. Pyruvate kinase is more
active in the fed state than in the fasting state.
- inhibited by:
Fates of Pyruvate
Pyruvate formed by glycolysis, is only the first stage in the complete
degradation of glucose.
First route: pyruvate is oxidized, with loss of its carboxyl group as
CO2, to yield acetyl group of acetyl-CoA by PDH. Acetyl group is
then oxidized to CO2 by TCA cycle.
Second route: pyruvate is reduced to lactate by lactate
dehydrogenase via lactic acid fermentation.
Third route: pyruvate catabolism leads to alcohol and CO2, under
hypoxic conditions, a process called alcohol fermentation.
Fructose:
a) is highly regulated
b) primarily metabolized by muscle
c) Uses GLUT5 transporter
d) is essential in fructosuria patients
42
Cytoplasm
cytoplasm
mitochondrial membrane
mitochondrial matrix
mitochondrial matrix
Glycolysis
PDH
TCA
ETC
Regulation of PDH
Allosteric
- Inhibition by ATP, acetyl-CoA and NADH.
TCA Cycle
Degradation of acetyl-CoA derived from carbohydrates, fatty acids
and amino acids.
- produces most of the CO2 generated in tissues (2).
- is the major source of NADH (3).
- allows excess energy to be used for fatty acid biosynthesis.
- provides precursors for many metabolites.
Most TCA cycle enzymes in the mitochondrial matrix; some in the
1
2
3
4
5
TCA Strategy
- Activated 2C fragment (acetyl-CoA) reacts with a 4C oxaloacetic acid, to
yield 6C citrate.
- In a series of seven reactions two carbons are released as CO2 (oxidative
decarboxylations), regenerating oxaloacetate.
- 2 carbons in, 2 carbons out = no net gain.
- Oxaloacetate must be present to "prime" the cycle.
- Four pairs of electrons are transferred during one turn of the cycle: three
pairs of electrons reducing three NAD+ to NADH, and one pair reducing
FAD to FADH2.
- One complete turn of the TCA cycle generates only one ATP (GTP) (in the
conversion of succinyl-CoA to succinate), three molecules of NADH and
one molecule of FADH2. Oxidation of one NADH leads to formation of
3 ATP, whereas oxidation of FADH2 yields 2 ATP.
-
Total 12 ATP generated by one round of TCA cycle from one acetyl residue. From
two acetyl residues, total 24 ATP generated.
TCA Cycle
Acetyl-CoA
citrate
synthase
2
malate
dehydrogenase
aconitase
TCA Cycle
isocitrate
dehydrogenase
fumarase
hydration
succinate
-ketoglutarate
dehydrogenase dehydrogenase
succinyl-CoA
synthetase
6
5
(ATP)
phosphorylation
4
oxidative decarboxylation
Regulation of the
TCA Cycle
Regulation of metabolite flow
from the PDH complex through
the TCA cycle in mammals.
The PDH complex is allosterically
inhibited when [ATP]/[ADP],
[NADH]/[NAD+], and [acetylCoA]/[CoA] ratios are high,
indicating an energy-sufficient
metabolic state. When these ratios
decrease, allosteric activation of
pyruvate oxidation results.
Anaplerotic Pathways
TCA cycle intermediates
Pyruvate carboxylase, malic enzyme, PEP-carboxykinase. Four rxns
Regeneration of NAD
Oxidative phosphorylation
Oxidative phosphorylation involves the reduction of O2 to H2O with
electrons donated by NADH and FADH2.
Photo-phosphorylation involves the oxidation of H2O to O2, with
NADP+ as electron acceptor.
Oxidative phosphorylation and photo-phosphorylation, both are
mechanistically similar both processes involve the flow of electrons.
Oxidative phosphorylation begins with the entry of electrons into the
respiratory chain.
NADH (carries electrons from catabolic reactions) and NADPH (supplies
electrons to anabolic reactions), are water-soluble electron carriers that
associate reversibly with dehydrogenases.
Flavoproteins
Iron-sulfur proteins
Cytochromes
Ubiquinone (UQ) or Coenzyme Q (CoQ)
Protein-bound copper
69
Flavoproteins
- The FAD or flavin mononucleotide (FMN) in the flavoproteins can
transfer either one- or two-electrons at a time.
- Electron transfer occurs because the flavoprotein has a higher
reduction potential than the compound oxidized.
- They accept hydrogen/electrons from NADH and donate it to the
cytochromes.
Fe3+ + e- Fe2+
Cytochromes
Cytochromes: Heme proteins (is a complex of protoporphyrin IX and
ferrous iron) that functions as electron carriers in respiration,
photosynthesis and other oxidation-reduction reactions.
- One-electron carriers (Fe3+ Fe2+).
- All but one (cytochrome c) are integral membrane proteins.
- Cytochrome c is a small and water-soluble protein.
What does it do?
A. Fe-S proteins
B. Flavin proteins
C. Cytochrome C
D.Cytrochome b
E. Coenzyme Q
(ubiquinone)
1. Integral
membrane
protein
2. Free to diffuse
Protein-bound copper
- Protein-bound copper participates in the last reaction of
the respiratory chain, the transfer of electrons to molecular
oxygen. It switches between the Cu+ and Cu2+ forms during
these electron transfers. Part of complex 4
Respiratory Chain
Four members of the respiratory chain are freely diffusible:
NADH, ubiquinone, cytochrome c, and molecular oxygen.
1 Complexes I and II catalyze electron transfer to ubiquinone from two
different electron donors: NADH (complex I) and succinate or
FADH2 (complex II).
2 Complex III carries electrons from reduced ubiquinone to cytochrome
c, and
3 Complex IV transferring electrons from cytochrome c to O2.
COMPONENTS OF THE
ELECTRON TRANSPORT CHAIN
COMPLEX I
COMPLEX III
COMPLEX IV
Complex I
(NADH to Ubiquinone)
- Complex I, also called NADH:ubiquinone oxidoreductase or
NADH
dehydrogenase, transfers electrons from NADH to ubiquinone, is a
large enzyme composed of 42 different polypeptide chains, including
FMN-containing flavoprotein and at least six iron-sulfur centers.
-no FAD?
- Electrons are transferred from NADH -> FMN -> ironsulfur -> ubiquinone.
Complex II
(Succinate to Ubiquinone)
Complex II, also called succinate dehydrogenase, is the only
membrane-bound enzyme in the TCA cycle.
Complex III
(Ubiquinone to Cytochrome c)
Complex III, also called cytochrome c oxidoreductase or cytochrome
bc1 complex, couples the transfer of electrons from ubiquinol (QH2) to
cytochrome c.
- It contains cytochrome b, an Fe-S protein, and cytochrome c1.
- Cytochrome c is a soluble protein. After its single heme accepts an
electron from Complex III, cytochrome c moves to Complex IV to
donate electron to a binuclear copper center.
INHIBITED BY ANTIMYCIN A
Complex IV
(Cytochrome c to O2)
This is the final step of the respiratory chain. Complex IV, also called
cytochrome oxidase, carries electrons from cytochrome c to molecular
oxygen, reducing it to H2O.
- Complex IV is a large enzyme (13 subunits; Mr 204,000) of the inner
mitochondrial membrane.
- Oxygen is tightly bound between heme a3 and copper, to be released
after its reduction to H2O by the transfer of four electrons.
ATP Synthase
O for Oligomycin
F1
FO
84
Proton Gradient
Oxidative phosphorylation proceeds in two steps:
1. Protons are pumped out of the mitochondrion: Proton pumping is
driven by the redox reactions in the respiratory chain, and creates a
electrochemical gradient across the inner mitochondrial membrane.
The inner membrane must be impermeable to protons to maintain
gradient.
Protons are admitted back into the mitochondrion, down their
concentration gradient, via proton channel: This process drives
ATP synthesis.
- four protons are pumped by the NADH-ubiquinone reductase complex
and another four by the ubiquinone-cytochrome c reductase complex.
- cytochrome oxidase removes four protons from the mitochondrial
matrix, translocating two to the inter-membranous space and other two
consuming in the reduction of O2, thus, four protons leads to the synthesis
of one ATP molecule.
- ATP is synthesized only when protons flow, and protons can flow
Respiratory Control
-
2,4-dinitrophenol:
A. Inhibits the flow of electrons at complex 4
B. lowers the basal metabolic rate
C. Inhibit the flow of electron at complex 2
D. decrease the H+ gradient across the inner
mitochondrial membrane
Mitochondrial DNA
Mutations in mitochondrial DNA can cause:
- Lebers hereditary optic neuropathy (LHON): blindness, caused by degeneration
of the optic nerve.
- Myoclonic epilepsy and ragged-red fiber disease (MERRF).
- Mitochondrial DNA has a higher mutation rate than nuclear DNA.
92
O2
HO2
HO2
O2
H2O2
H+
Hydroxyl radical:
H2O2 + Fe2+ + H+ HO + Fe3+ + H2O
SOD
H2O2 + O2
Catalase
2H2O + O2
Peroxidases
Glutathione peroxidase (2GSH + H2O2 GSSG + 2H2O)
Bilirubin
Uric acid
Ascorbate (Vitamin C)
Estrogens
Carbohydrate Metabolism
Most important function of carbohydrate metabolism is: maintain blood
glucose level at all times.
- Brain alone consumes ~ 120 g of glucose / day. Therefore, a blood glucose
level of 4.0 to 5.5 mmol/liter (70 to 100 mg/dL) must be maintained at all
times.
-
Liver produces 10 times more glucose than kidney, because the liver is so
much larger than a kidney.
- Brain alone requires ~ 120 g of glucose each day this is more than
half of all the glucose stored as glycogen in liver and muscle.
- Liver is the major gluconeogenic organ of the body.
- Muscle does NOT contribute to gluconeogenesis, because
Gluconeogenesis
Gluconeogenesis: synthesis of carbohydrate (such as glucose)
from non-carbohydrates such as oxaloacetate or pyruvate
(Synthesis of one molecule of glucose from two molecules of pyruvate
requires energy, equivalent to 6 molecules of ATP)
Gluconeogenesis
Gluconeogenesis and glycolysis are NOT identical pathways,
running in opposite directions. Reverse of glycolysis except
the irreversible (kinase) reactions.
Three reactions of glycolysis are irreversible and cannot be used in
gluconeogenesis:
1. Hexokinase/
Glucokinase
2. PFK-1
3. Pyruvate Kinase
A. PEP carboxykinase
B. Fructose -1,6bisphosphatase
C. Pyruvate carboxylase
D. Glucose-6phosphatase
Glycolysis
Gluconeogenesis
[10]
[8]
Glycolysis and
Gluconeogenesis
Irreversible reactions # [1], [8],
and [10] Know the enzymes.
7
6
5
4
3
2
[1]
Notice the use of an ATP and a GTP
the liver.
- Glucose can thus leave the liver and enter the blood.
Note: glucose-6 phosphatase is involved in both gluconeogenesis and
glycogenolysis.
Fructose-2, 6-bisphosphate:
A. Stimulates glycolysis
B. Is inhibited by PFK1
C. Is inhibited by PFK2
D. Favors gluconeogenesis
E. Stimulates Fructose-1,6-bisphosphatase
Activates glucose
breakdown
Gluconeogenic Substrates
- Lactate (Pyruvate) Cori cycle. Lactate is released into the blood by
muscle, and by cells that lack mitochondria, such as RBCs and taken
to the liver to be used for gluconeogenesis
- Glucogenic amino acids (all, except leucine and lysine).
- Glycerol (from the breakdown of triglycerides in adipose tissue),
-
Cori Cycle
Cori Cycle: Shuttling of glucose and lactate between muscle and liver
during physical exercise.
Regulation of Gluconeogenesis
Hormones are important for the regulation of gluconeogenesis:
Regulation of Gluconeogenesis
Glucagon and insulin secreted by pancreatic and cells,
respectively. The release of glucagon and insulin is inhibited
by somatostatin, which is secreted by the pancreatic
delta cells.
Gluconeogenesis
-
A.Glycolysis
B.Gluconeogenesis
C.Fatty Acid Oxidation
D.A & B
Why Glycogen?
- Fat cannot be as rapidly mobilized in muscle.
-Most people have more muscle than liver, the total amount of muscle
glycogen exceeds that in the liver.
There is more glycogen per gram of tissue weight in the liver (it is more
concentrated in the liver)
Glycogen Structure
6
6
5
5
1
4
3
5
1
4
3
Glycogen Synthesis
Glycogen is readily synthesized from glucose.
(Glucose is a most abundant monosaccharide in dietary carbohydrate)
For glycogen synthesis:
- Primer required to initiate synthesis.
- The protein glycogenin (Mr 37 kDa) serves as the primer,
or that primes the synthesis of new glycogen chains.
Glycogen Synthesis
Glycogen Synthesis
Glycogen synthase is the key regulatory enzyme for glycogen synthesis;
creates the -1,4 glycosidic bonds in glycogen by transferring the
glucose residue from UDP-glucose to non-reducing end of the
glycogen molecule.
Glycogen synthase cannot form the -1,6 glycosidic bonds at the branch
points of glycogen. Branching requires a glycogen-branching
enzyme, called transglycosylase or glycosyl-transferase, that breaks
an -1,4 bond and forms an -1,6 bond .
Glycosidic bonds: are covalent bonds, formed between two
monosaccharide molecules by means of a dehydration reaction.
Glycogen phosphorylase
Hexokinase
Glucose-6-phosphatase
Phosphoglucomutase
All of the above are found in muscle
cAMP
C C
R R
Inactive
C C
Active
Kinase A
Protein Kinase A
Ca++ (muscle)
ATP
OH
Inactive
phosphorylase kinase
Phosphoprotein
phosphatase
AMP(muscle)
O-P
Active
Phosphorylase kinase
Glucose
2 ATP
OH OH
OP OP
Phosphorylase b
Phosphorylase a
Phosphoprotein
phosphatase
Glycogen
Glucose-1P
128
NADPH:
A. Maintains an oxidative environment of the
cytosol
B. Provides electrons for oxidative
phosphorylation
C. Is a coenzyme for fatty acid synthesis
D. Is mainly found in the mitochondrion
Which Tissues?
- all tissues:
2 phases
Oxidative Phase: oxidative branch of the PPP is concerned
with the synthesis of NADPH. Glucose-6-phosphate
dehydrogenase catalyzes the rate-limiting step. The reaction
is irreversible, maintains a high NADPH / NADP+ ratio.
Balance Sheet
Summary of the flow of 15 C atoms through Pentose Phosphate
Pathway reactions by which 5-C sugars are converted to 3-C and 6C sugars.
C5 + C5 C3 + C7
C3 + C7 C6 + C4
C5 + C4 C6 + C3
3 C5 2 C6 + C3
(Transketolase)
(Transaldolase)
(Transketolase)
(Overall)
If energy is also needed, 5C products are recycled to glucose-6phosphate for glycolysis and repeat PPP reactions (e.g. RBCs).
140
Glutathione
- Glutathione (GSH, tripeptide; Glu-Cys-Gly), functions as a
reducing agent.
- Glutathione Peroxidase (GPx) catalyzes degradation of
hydroperoxides by reduction, as two glutathione molecules
(represented as GSH) are oxidized to a disulfide (GSSG).
Only the reduced form of glutathione (GSH) is an antioxidant.
Therefore, the dimeric, oxidized form (GSSG) has to be reduced back
by the enzyme glutathione-reductase (GR). Can be faulty
(GR depends on NADPH from the pentose phosphate pathway)
2 GSH + ROOH
GPx
GSSG + NADPH + H+
GR
2 GSH + NADP+
NADPH + H+
GS-SG
NADP+
2 GSH
HSProteinHS
Protein
Active enzyme
Denatured enzyme
1.Liver
2.Adipose
3.Muscle, and
4.Brain.
Insulin
Glucagon
Glucagon: is a polypeptide hormone secreted by
the -cells of the pancreas that stimulates the
glucose-producing pathways of the liver. It is
released in response to hypoglycemia (decreased
blood glucose Level, < 40 mg/dl). Hypoglycemia is
a medical emergency.
Glucagon raises the blood glucose level.
Glucagon promotes the release of glucose from the
liver.
Glucagon stimulates glycogenolysis and gluconeogenesis
and inhibits glycolysis. in the liver
a) 55mmol/liter
b) 90mg/dL
c) 5mg/dL
d) 0.5mmol/liter
F-2,6 bisphosphatase
Inhibition of:
(loss of fructose-2,6-bisphosphate)
PFK-2
Phosphorylase kinase
Pyruvate kinase
Glycogen phosphorylase
Glycogen synthase
Hormone-sensitive lipase
Re-feeding
The levels of glycolytic enzymes in the liver are very low, and
patients severely starved show profound carbohydrate intolerance.
Therefore, re-feeding should be started slowly.
Activation of:
- F-2,6 bisphosphatase
- PFK2
- Phosphorylase kinase
- Pyruvate kinase
- Glycogen phosphorylase
- Glycogen synthase
- Hormone-sensitive lipase
and older individuals. It is more common than type 1, is less severe. The
pancreatic cells are intact, and the plasma level of insulin may be
normal, reduced, or elevated. The problem may be either reduced insulin
secretion or insulin-resistance of the target tissues, or a combination of
both.
- Patients with type 2 diabetes are not afflicted by ketoacidosis b/c they
have insulin to inhibit the breakdown of Fat to acetyl CoA, Acetyl-CoA
doesnt accumulate, so it doesnt get converted to ketones. It is
characterized by excessive glucosuria (glucose in the urine) with
osmotic diuresis (high glucose levels in the urine leads to concentrated
urine, which pulls water into the urine to dilute it) . If the patient
forgets to drink, the resulting dehydration can become sufficiently
severe to affect the central nervous system.
- The over-treatment of diabetes with insulin or oral anti-diabetic drugs
leads to hypoglycemic shock.