REVIEW BIOCHEMISTRY

LECTURE 19- LIPIDS & LIPOPROTEINS

Review of Key Concepts

Storage lipids are :
1.
2.
3.
4.
5.

Simplest lipids present in adipocytes

Nonpolar, neutral and hydrophobic
3 fatty acids esterified to a glycerol
Serves as an alternate energy source
Mobilized under glucagon; stored under insulin action

Phospholipids are:
1.
2.
3.
4.

Structural lipids present in cell membranes

Have a hydrophilic head with negative charge
Hydrophobic tail composed of 1 or 2 fatty acids
Phosphoglycerides, sphingolipids

Glycolipids:
1.
2.
3.
4.

Head is composed of simple/complex sugar

May be charge bearing
Have ceramide backbone
Responsible for ABO blood grouping

Review
PUFAs are:
1.
2.
3.
4.
5.

Present in plant fats

Have multiple double bonds (cis) in the C skeleton
Have low melting temperature; liquid in RT
Good for health
Omega fatty acids are PUFAs

Saturated fatty acids are:

1.
2.
3.
4.

Present in animal fats

Have no double bonds in the C skeleton
Have high melting temperature; solid at RT
Not so good for health

Trans fats:
1. Have trans double bonds in the C skeleton
2. Produced from PUFAs when exposed to high heat/ hydrogenation
3. Unhealthy

Review
Chylomicrons are:
1.
2.
3.
4.

Made in the small intestine

Transport dietary fats via blood for storage or usage
Composed of ApoB48, ApoC, ApoE
Carries principally dietary triacylglycerol (TG)

VLDLs & LDLs are:

1.
2.
3.
4.
5.

VLDL produced in liver; LDLs produced from VLDLs

VLDLs carry TG made in liver; LDLs carry cholesterol
Composed of Apo B100, ApoC, ApoE
High levels in blood is a risk factor for MI
High trans fat & saturated fat intake increase levels of VLDL/LDLs

HDLs are:
1. Good cholesterol; induces reverse transport of cholesterol
2. PUFAs increase HDL production
3. Contains ApoA1

LECTURE 20
FATTY ACID & CHOLESTEROL SYNTHESIS

Review
Palmitic acid is the primary FA synthesized in the
cytoplasm; modified into other types of FAs by
elongation/desaturation
Phase I: Formation of acetyl CoA to malonyl CoA;
Enzyme: Acetyl CoA carboxylase (rate limiting
enzyme); biotin
dependent; citrate acts as a + regulator;
activated by
dephosphorylation (Insulin); inactivated by
glucagon(phosphorylation)
Phase II: 7 repeated cycles of condensation
reduction, dehydration & reduction; the enzymes
involved belong to the FAS complex; requires 1
acetyl CoA, 7 malonyl CoA, 14 NADPH
Citrate shuttle makes mit. acetyl CoA available as
citrate in the cytoplasm for FA synthesis; Malate
shuttle brings the oxaloacetate back to the Mit.

Review
Liver cells synthesize 50% of endogenous
cholesterol
Synthesized from acetyl CoA & acetoacetyl
CoA
Rate limiting enzyme is HMG-CoA reductase
Activated by Insulin; inactivated by glucagon
Inhibited by statin (competitive inhibitor)
Statin suppresses intracellular cholesterol
synthesis
Statin increases LDL clearance from blood;
long term use lowers prenyl protein levels in
the cell, affects ETC
High intracellular cholesterol degrades HMGCoA-reductase; inhibits cholesterol
biosynthesis
Low intracellular cholesterol activates SREBP

LECTURE 21 FATTY ACID OXIDATION

Review

Beta oxidation involves successive removal of 2C

from COOH end of FA as acetyl CoA; 1 FADH2 & 1
NADH is released per cycle; the last 2C are left as
acetyl CoA
Formation of each acetyl CoA requires 4 reactions;
dehydrogenation, hydration, dehydrogenation &
reaction with CoASH
Acetyl CoA feeds into the TCA cycle; FADH2 & NADH
feeds into ETC
Last 3C of odd chain FAs produce a propionyl CoA
during oxidation; it enters TCA cycle as succinyl CoA
Production of high energy phosphate bonds (ATPs/
GTPs) by complete oxidation of FAs depends on
their chain lengths
Over oxidation of FAs leads to ketosis

Review

FAs oxidation occurs in the mitochondria. Produces acetyl

CoA for feeding into TCA; the process of entry of FAs into
the mit. depends on their chain lengths
Long chain & very long chain FAs require carnitine shuttle to
enter into the Mit.
Steps of entry: Activation of FA to acyl CoA (2ATPs);
attachment with carnitine; entry into inter membrane space
(CPT-I mediated); entry into matrix (translocase mediated);
release of carnitine via CPT-II in mit. matrix
FA oxidation is upregulated by glucagon; inhibited by
insulin; higher cyt. malonyl CoA conc. inhibits CPT- I
Primary systemic carnitine deficiency: Very high urine
carnitine; very low plasma carnitine levels
CPT-I deficiency: Very high plasma carnitine; hypoketotic
hypoglycemia

CPT-II deficiency: Low plasma carnitine; High acylated

carnitine in blood
Synthesis & Oxidation of FAs cannot occur simultaneously
When new FAs are synthesized; CPT-I is turned off such they

LECTURE 22
ENZYMES OF DIAGNOSTIC IMPORTANCE

Review
Specific enzymes often are used as diagnostic markers of specific
clinical conditions
Different clinical conditions may affect the activity, concentration &
localization of the marker enzymes
When using enzymes as diagnostic markers we analyze
More than one marker enzymes
Different isoenzymes of a specific marker enzyme
Change in activity, conc., localization of the enzyme(s) at cellular
levels and enzyme clearance pattern
Routine photometric analysis is done to measure enzyme activity
proportional to enzyme concentration in labs
We can quantify the enzyme(s) using immunological techniques (like
ELISA)
Direct ELISA: Simplest process; uses a labeled 1Ab for detection &
quantification
Indirect ELISA: Uses 2 Ab. 1Ab is unlabeled; 2Ab is labeled. Used
for measuring endogenous antibody titers for detection of infections

Review
a. Liver diseases: High ALT & AST (markers of hepatocellular
damage); ALT (a more specific marker) is higher than AST;
AST/ALT ratio<<<1
b. Alcohol induced liver damage: High AST when compared to ALT
; High GGT; AST/ALT ration >>1
c. Cholestasis : Very high ALP levels; significantly high AST and
ALT levels; High bilirubin in blood jaundice like symptoms;
change in color of stool /urine
d. Bone diseases: High ALP, bone pain, hypercalcemia, may show
Vit D def.
e. Pancreatitis: High Amylase and lipase; inability to digest fat;
steatorrhoea
f. Pancreatic Pseudocyst:
Very High Amylase levels; similar
symptoms as pancreatitis
g.MI: High Myoglobin immediately after the incident
Presence of CK2,
Troponin T/I (specific markers of
cardiomyopathy) after 2-6 hr of the incident; High levels of CK-2,
total CK, Troponin I/T 24 hr after the incident

LECTURE 23
ASMA PROTEINS & ASSOCIATED DISORDERS

Review
Blood serum is plasma without the coagulation factors
Majority of the plasma/ serum proteins are synthesized in the
liver except
gamma globulins
Plasma/ serum hypo/hyperproteinemia has clinical significance
Acute phase reactants (APR): Serum/plasma proteins the levels
of which are affected by inflammatory conditions
+ APRs: Concentration increases (majority of plasma proteins)
-APRS: Concentration decreases (albumin, transferrin)
Serum/ plasma electrophoresis: Separates serum/plasma
proteins according to MW & negative charge.
Albumin (smallest & most negative): migrates fastest; gets
closest to the anode (+)
Gamma globulins (largest & least negative): Moves slowly,
remains closest to the cathode(-)

Review
Albumin: Major plasma protein, small, extremely hydrophilic;
highly negative; is not eliminated by kidneys; strongest buffer in
blood
Albumin is the major contributor of colloidal osmotic pressure or
oncotic pressure; transports copper via blood
Hypoalbuminemia (caused by liver/ kidney diseases/
malnutrition) leads to edema (cause: low oncotic pressure)
Globulins: Higher MW, low negative charge; subtypes are 1, 2,
,
1 antitrypsin: 1 globulin; inactivates proteases at cellular
levels; deficiency causes emphysema due to elastolytic damage
of lung alveoli by active elastase in the lung tissue
Cigarette smoke inactivates 1 antitrypsin in plasma;
contributes to emphysema in chain smokers

Review
Haptoglobin: 2 globulin, synthesized by the liver; carries
intravascular Hb-dimers (consequence of intravascular
hemolysis) to the liver/ macrophages for bilirubin synthesis. Free
haptoglobin levels are very low during severe intravascular
hemolysis
Hemoglobinuria: Hb in urine: caused by high intravascular
hemolysis/low haptoglobin synthesis by liver
Ceruloplasmin: 2 globulin, synthesized by the liver; principle
copper transporter present in blood; promotes iron transport via
blood
Apoceruloplasmin (precursor) when binds to 6 copper atoms
forms active ceruloplasmin in hepatocytes. Catalyzed by ATP7B
Wilsons disease: Defective ATP7B; low ceruloplasmin, excess
copper in liver; Cu leaks out in blood; organ damage due to
abnormal copper deposits
Menkes syndrome: Defective ATP7A; low ceruloplasmin &
copper in blood; lack of copper at tissue levels leads to organ

Review
Multiple Myeloma (MM): common form of hematological cancer;
caused by abnormal proliferation of a single clone of plasma
cells; Single class of Ab produced in excess; 5 main types
Symptoms: Bone pain; osteolytic lesions (X ray); low A:G ratio
in serum; M spike detectable in plasma protein electrophoresis;
Bence Jones protein present in urine electrophoresis (light chain
MM); hypercalcemia; kidney defect, anemia , thrombocytopenia
Tests: Bone X ray; serum/ urine electrophoresis with immuno
fixation (for identifying the specific type of MM); bone marrow
biopsy; genetic tests
Waldenstroms macroglobulinemia: Clonal disease of B
lymphocytes; characterized by abnormal production of IgM; rare
type of hematological cancer
Symptoms: Hyperviscosity syndrome (cardiac issues); anemia,
thrombocytopenia; cryoglobulinemia (causes tingling of
hands/feet in cold)

LECTURE 24
HEMOSTASIS & THROMBOSIS

Revie
2 physiological regulators
w of coagulation

Protein C: Produced by liver & activated by thrombin;

blocks formation of tenase, prothrombinase complexes
by inactivation Va, VIIIa; turns off thrombin production
& hemostasis

Antithrombin III: Neutralizes available thrombin; turns

off factors required in secondary hemostasis turning
off thrombin production

Heparin : Mimics antithrombin III; used as short term

coagulant
Warfarin : Long term coagulant; antagonist of Vit K;
reduces available Vit K levels in the body; slows down
coagulation
Vit K: Cofactor of the liver enzyme GGC which
carboxylates & activates factors II, VII, IX, X; deficiency
slows down coagulation

Revie
Hemostasis: Normal process of clot formation in
w
response to a vascular injury

Thrombosis: Formation of clot in intact blood vessel;

pathological

Steps of hemostasis
a. Primary: Formation of platelet plug; temporarily
stops blood loss
b. Secondary hemostasis: Formation of the fibrin
meshwork around
the plug; stops blood loss permanently
c. Tertiary: Dissolution of fibrin meshwork
Primary hemostasis
TxA2: Activates platelets for aggregation; Aspirin blocks
TxA2 production from platelets; used as blood thinner
vWF + factor VIII (from damaged endothelium): Initiates
platelet plug formation; TxA2, thrombin, MLCK promotes it

Revie
Secondary hemostasis: w

Extrinsic: Short process; forms small amounts of Xa;

involves thromboplastin, factor VII & factor X; can form
a small clot; Ca+2 dependent

Intrinsic: longer process; dependent on contact

activation; involves activation of factors XII, XI, IX, VIII
and X; activates large amount of Xa; involves tenase
complex formation (Ca+2 dependent)

Common pathway: Xa enters common pathway; forms

prothrombinase complex with Va (Ca+2 dependent);
activates prothrombin to thrombin; forms insoluble
fibrin polymers from fibrinogen (via soluble fibrin
monomers), involves factor XIIIa

Tertiary hemostasis:
Fibrinolysis is the main event; tPA activates plasminogen
to plasmin which degrades fibrin to fibrin degradation

LECTURE 25
BIOCHEMICAL ENDOCRINOLOGY

Review
Hormones: Chemicals secreted by endocrine glands into

blood;
acts on remote target organs; receptor mediated action
Protein/peptide hormones: Gene products; produced in
inactive form; activated by post-translational modifications in
ER/ golgi; stored in secretory vesicles in cytoplasm; released
by exocytosis into blood
Post Pit. Hormones: Oxytocin & Vasopressin; Synthesized in
hypothalamus; stored & released from post. Pit.
Diabetes insipidus: Low ADH levels (central DI); insensitivity
of kidneys to ADH (nephrogenic DI); symptoms are polyuria,
polydipsia
Insulin: Preproinsulin, proinsulin, active insulin + C peptide
Diabetes type 1A: Autoimmune destruction of beta cells; low
insulin and c peptide levels
Diabetes Type 1B: Non-responsive beta cells; no insulin or Cpeptide

Review
Thyroid hormones: T3 (active form) and T4; made from
iodinated tyrosine residues in follicular epithelial cells of
thyroid gland; can diffuse through cell membrane; act via
cytoplasmic /nuclear receptors
Thyroglobulin supplies tyrosine for T3/T4 synthesis; iodine is
supplied from diet.
Transporters involved: NIS, Pendrin
Enzymes involved: TPO; Dual oxidases
Congenital hypothyroidism: Low T3/T4 levels; High TSH;
goiter; iodine deficiency is the common non genetic cause;
defective Tg, NIS, TPO, pendrin can be other genetic causes
Epinephrine/ norepinephrine: Synthesized in adrenal medulla
in response to sympathetic stimulation; is not a gene product;
made from tyrosine
Pheochromocytoma: Adrenal medullary tumors;

a.
b.

Review

All Steroid hormones are made from cholesterol.

Regulatory steps and enzymes:
Transfer of cholesterol from cytoplasm to mitochondria - STAR
Conversion to pregnenelone in the mit.- CYP11A1

. Adrenal cortex:
a. Glomerulosa: lacks 17-hydroxylase, 11-hydroxylase.
Contains aldosterone
synthase; produces only aldosterone
b. Fasiculata: Has 17-hydroxylase, 11-hydroxylase;
lacks aldosterone synthase; produces cortisol & small
amounts of adrenal steroids
c. Reticularis: Has 17-hydroxylase, 11-hydroxylase ;
lacks 21 hydroxylase & aldosterone synthase; produces
adrenal steroids(DHEA, androstenedione)
CAH: Deficiency of 21 hydroxylase in adrenal cortical cells;
overproduction of adrenal steroids; Classical and non classical
types
Testosterone: Produce from cholesterol in Leydig cells