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Vitamins:
1. Explain the role of ascorbic acid in post translational modification. Pon May
2007
a. The biosynthesis of a protein or a polypeptide in a living cell is referred to as
translation. The proteins synthesized in translation are, as such, not functional.
Many changes take place in the polypeptides after the initiation of their synthesis
or, after the protein synthesis is completed. These modifications include protein
folding, trimming by proteolytic degradation, intein splicing and covalent changes
which are collectively known as modifications. Covalent modifications include
phosphorylation, hydroxylation and glycosylation.
b. Vit C plays a role in posttranslational hydroxylation of proline and lysine
c. Hydroxylation : During the formation of collagen, the amino acids proline and lysine
are respectively converted to hydroxyproline and hydroxylysine. This hydroxylation
occurs in the endoplasmic reticulum and requires vitamin C.
2. FOLATE TRAP:
1. In vitamin B12 deficiency, increased folate levels are observed in plasma. The activity
of the enzyme homocysteine methyltransferase is low in B12 deficiency. As a result,
the only major pathway for the conversion of N5-methyl THF to tetrahydrofolate is
blocked and body THF pool is reduced. Essentially almost the entire body folate
becomes trapped as N5-methyl THF. This is known as folate trap or methyl frap.
2. ln this manner, B12 deficiency results in decreased folate coenzymes that leads to
reduced nucleotide and DNA synthesis. Although the tissue folate levels are adequate
or high, there is a functional folate deficiency due to the lack of THF pool. The outcome
is the development of megaloblastic anemia.
3. Administration of the amino acid methionine has been shown to partially correct the
symptoms of B12 deficiencies. This is due to the fact that the formation of N5-methyl
THF is inhibited by S-adenosylmethionine. A combined therapy of vitamin B12 and
folic acid is generally employed to treat the patients with megaloblastic anemia.
3. Schillings test:
1. The test is for assessing weather B12 absorption from the gut is normal. It is done in
two stages.
2. Stage I: Radioactive labelled (Cobalt-60) vitamin B12, one microgram is given orally.
Simultaneously an intramuscular injection of unlabeled vitamin B12 is given to
saturate tissues with normal vitamin B12 and radioactive vitamin B12 will not bind to
body tissues. Therefore, the entire absorbed radioactivity will pass into the urine. The
patients urine is then collected over the next 24 hours to assess the absorption.
3. In patients with pernicious anemia or with deficiency due to impaired absorption, less
than 5% of the radioactivity is detected in urine.
4. Stage II: If an abnormality is found, the test is repeated, with radioactive vitamin plus
intrinsic factor given orally, and urine is collected for 24 hours. In pernicious anemia,
there is lack of intrinsic factor, so that the first test is abnormal; but the second test is
normal.
4. The elevated excretion of formimino glutamate (FIGLU) occurs in which
condition? Give the reaction blocked. Pon Nov 2006
Ans: In the metabolism of the amino acid histidine there is folic acid dependent step at the
point where formimino-glutamic acid (Figlu) is converted to glutamic acid. In folic acid
deficient patients, this reaction cannot be carried out, as a result, figlu accumulates in
the blood and excreted in urine. Figlu excretion in urine is an index of folic acid deficiency.
When a loading dose of histidine is given, the excretion of Figlu in urine is increased
further (Histidine loading test).

5. Why does the dietary requirement of pyridoxin increase with high protein diet?
Pon May 2010
a. Pyridoxine acts in many reactions of amino acid metabolism. Synthesis of certain
specialized products such as serotonin, histamine & niacin.
b. There is evidence that requirement of Vitamin B6 is increased due to increased
dietary protein intake, as it is involved as coenzyme in many metabolic reactions of
amino acid metabolism.
c. Pyridoxal phosphate participates in reactions like transamination, decarboxylation,
deamination, transsulfuration & condensation reactions of aminoacids.
d. Transamination:
i. Eg. transaminase converts aminoacids to keto acids with PPL acting as
coenzyme. The keto acids enter the citric acid cycle and get oxidized to
generate energy.
Alanine + Alpha keto glutarate Pyruvate + Glutamic acid (Enzyme Alanine
transaminase).
e. Decarboxylation: examples:
i. Tryptophan is first converted to 5-hydroxy tryptophan and then
decarboxylated to serotonin in the presence of PLP.
ii. Histidine is converted to histamine in the presence of PLP.
iii. Glutamate is converted to Gama amino butyric acid by similar
decarboxylase reaction with PLP.
iv. Catecholamines are synthesized from tyrosine by decarboxylase and PLP.
f. Sulfur Containing Amino Acids (Transulfuration):
i. PLP plays an important role in methionine and cysteine metabolism:
ii. Homocysteine + Serine Cystathionine. (Enzyme Cystathionine synthase)
iii. Cystathionine Homoserine + Cysteine (Enzyme Cystathionase)
g. In heme synthesis: PLP is required for the condensation of succinyl CoA and
glycine to form delta amino levulinic acid (ALA). In B6 deficiency, anemia may be
seen.
h. In deamination reactions
i. Dehydratase enzyme converts serine to pyruvate in the presence of PLP.
ii. Pyridoxal phosphate is required for the synthesis of niacin from
tryptophan i.e: Coenzyme for kynureninase. This is instance in which one
vitamin synthesizes another vitamin.
iii. Hydroxymethyltransferase converts serine to a-ketoglutarate with PLP.
i. Other important functions of PLP:
i. Synthesis of sphingolipid and myelin formation.
ii. Absorption of amino acid from intestines
iii. Formation of Coenzyme A
iv. Boosting immune functions
v. Preventing urinary stone formation
vi. Utilization of unsaturated fatty acids.
6. Which enzyme in RBC is measured in thiamine deficiency? Name the other
enzymes that require thiamine for their activity. Pon Dec 2002
1. In thiamine deficiency, blood thiamine is reduced, but pyruvate, alpha ketoglutarate
and lactate are increased.
2. Transketolase: The second group of enzymes that use TPP as co-enzyme are the
transketolases, in the hexose monophosphate shunt pathway of glucose.

3. Erythrocyte transketolase activity is reduced with accumulation of pentose sugars;

this is the earliest manifestation seen even before clinical disturbances. Measurement
of RBC transketolase activity is a reliable diagnostic test to assess thiamine deficiency.
7. Beriberi:
1. This is a severe thiamine-deficiency syndrome found in areas where polished rice is the
major component of the diet. It is characterised by the following manifestations:
2. CVS:
a. These include palpitation, dyspnoea, cardiac hypertrophy and dilatation, which
may progress to congestive cardiac failure.
3. Neurological manifestations:
a. These are predominantly those of ascending, symmetrical, peripheral
polyneuritis.
b. Polyneuritis common in chronic alcoholics. Alcohol utilization needs large doses
of thiamine. Alcohol inhibits intestinal absorption of thiamine, leading to
thiamine deficiency. Polyneuritis may also be associated with pregnancy and old
age. Thiamine deficiency may cause impairment of conversion of pyruvate to
acetyl CoA. This results in increased plasma concentration of pyruvate and
lactate, leading to lactic acidosis.
c. Wernickes encephalopathy: This is seen primarily in association with chronic
alcoholism and is due to dietary insufficiency or impaired intestinal absorption
of the vitamin. It is characterized by ophthalmoplegia, nystagmus, cerebellar
ataxia.
d. Some alcoholics develop Wernicke- Korsakoff psychosis syndrom characterized
by apathy, loss of memory, ataxia, and a rhythmic to-and-fro motion of the
eyeballs (nystagmus).
e. The neurologic consequences of Wernicke's syndrome are treatable with
thiamine supplementation.
4. GI symptoms:
a. Amongst these, anorexia is an early symptom. There may be gastric atony, with
diminished gastric motility and nausea; fever and vomiting occur in advanced
stages.
5. Dry beriberi: When it is not associated with oedema.
6. Wet beriberi: Oedema is associated. It is probably in part to congestive cardiac failure
and in part to protein malnutrition (Low plasma albumin).
7. Infantile beriberi: It occurs in infants born to mothers suffering from thiamine
deficiency. Signs of infantile beriberi include restlessness and sleeplessness
tachycardia, vomiting, convulsions, and, if not treated, death.
8. ONE-CARBON METABOLISM
1. One-carbon (1C) groups play a pivotal role in donating carbon atoms for synthesis of
different types of compounds.
2. One carbon groups are:
a. Formyl group
b. Formimino group
c. Methanyl group
d. Hydroxymethyl group
e. Mrthylene group
f. Methyl group

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3. Except mehyl group others are carried by tetrahydrofolic acid. They are contributed by
by amino ac-ids
4. Co2 is also a one carbon unit as it participates in carboxylation but not accepted by
many as a one arbon unit.
Formation of one carbon units:
1. The formate released from glycine and tryptophan metabolism combines with THF
to form N10- formyl THF.
2. Histidine contributes formimino fragment to produce N5-formimino THF
3. When serine is converted into glycine N5-N10 methylene THF is formed.
4. Choline and betain contribute to the formation of N5-methyl THF. Choline and
betaine are donors of hydroxy methyl groups. As serine is converted to choline, 3
one-carbon units are used up. During the conversion of choline to glycine, these
methyl groups are recovered. Hence, this pathway is called the "salvage pathway"
for one-carbon units.

Utilization of one carbon moiety:

One carbon units are used to synthesise the following compounds:
1. C2 of purine
2. Formylation of Methionyl t RNA
3. C8 of purine
4. Glycine
5. Pyrimidine nucleotide
Role of methionine and B12:
1. Active methionine is a methyl donor; after release of methyl group it becomes
homocysteine. Reconversion of homocysteine to methionine using N5-methyl
THF is a reaction which is catalyzed by a B12-containing methyltransferase
2. B12 coenzyme accepts methyl group from methyl THFA to form methyl
cobalamin. In B12 deficiency this transfer cannot occur and hence THFA (Folic
acid) cannot be regenerated. So folic acid deficiency coexists with B12
deficiency a condition called folate trap.
3. Most of the one carbon moities are metabolically interconvertible and
catalysed by an NADP-dependant hydroxymethyl dehydrogenases
9. Symptoms of Vit.K deficiency are met with more due to liver dysfunction than
due to lack of the vitamin. Explain why this is so. Pon apr 2002
1. Causes for Deficiency of Vitamin K
a. In normal adults dietary deficiency seldom occurs since the intestinal bacterial
synthesis is sufficient to meet the needs of the body. However deficiency can
occur in conditions of malabsorption of lipids. This can result from obstructive
jaundice, chronic pancreatitis, sprue, etc.
b. Prolonged antibiotic therapy and gastrointestinal infections with diarrhoea will
destroy the bacterial flora and can also lead to vitamin K deficiency.
10. How many mg of tryptophan are required to form 1 mg of niacin? Pon May
2009

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a. NAD can also be synthesized from the dietary amino acid tryptophan via the
kynurenine pathway. Tryptophan produces quinolinate which then forms nicotinate
mononucleotide and, ultimately, NAD+ and NADP+.
b. 60 mg of tryptophan would yield 1 mg of Niacin.

Minerals
1. Name the enzyme in heme biosynthetic pathway that require Zinc. Which metal
can inhibit this enzyme? Pon May 2007
a. Porphyrin synthesis occurs by condensation of four molecules of porphobilinogen
b. Two molecules of 6-aminolevulinate condense to form porphobilinogen (PBG) in the
cytosol.
c. This reaction is catalysed by a Zn-containing enzyme ALA dehydralase. lt is
sensitive to inhibition by heavy metals such as lead.
2. Can copper deficiency cuase anemia? How? Pon April 2002
a. Copper is necessary for iron absorption because Ceruloplasmin( a copper
containing protein) promotes iron absorption.
b. Ceruloplasmin is necessary for the incorporation of iron into transferrin.
Ceruloplasmin serves as ferroxidase and is involved in the conversion of iron from
Fe2+ to Fe3+ in which form iron (transferrin) is transported in plasma.
c. Copper is also necessary for the synthesis of hemoglobin as Cu, a constituent of
ALA synthase, is needed for heme synthesis.
d. Copper deficiency produces microcytic hypochromic anaemia, due to impairment of
erythropoiesis and decreases in erythrocyte survival time, which cannot be
corrected by administration of iron.
3. Role of selenium in the body? Pon May 2009; Mention the selenium requiring
enzymes in the body. Pon May 2013
1. Selenium intake depends on the nature of the soil in which food crops are grown.
Requirement is 50-100 microgram/day. Normal serum level is 50-100 microgram/dl.
2. Biochemical functions:
a. Selenium containing enzymes:
a. Glutathione peroxidase (GP) is the important selenium containing enzyme.
RBC contains good quantity of glutathione peroxidase. This enzyme protects
the cells against the damage caused by H2O2
b. Thyroxin is converted to T3 by 5'-de-iodinase, which is a selenium
containing enzyme. In Se deficiency, this enzyme becomes less active,
leading to hypothyroidism.
b. Selenium concentration in testis is the highest in adult tissue. It is necessary
for normal development of spermatozoa. It is concentrated in the mid-piece
of spermatozoa as a specific seleno-protein in mitochondria.
c. Se is involved in maintaining structural integrity of biological membranes
d. Se prevents lipid peroxidation and protects the cells against the free
radicals, including superoxide.
e. Se binds with certain heavy metals (Hg, Cd) and protects the body from
their toxic effects.
f. Selenium acts as a nonspecific intracellular anti-oxidant. This action of Se is
complementary to vitamin E. Availability of vitamin E reduces the selenium
requirement. In Se deficiency, tissue vitamin E content is depleted.
Selenium, along with vitamin E, prevents the development of hepatic
necrosis and muscular dystrophy
3. The UGA codon is acting as the codon for direct insertion of seleno-cysteine into
selenium containing enzymes. Seleno-cysteine is directly incorporated into the
protein during biosynthesis. So, seleno-cysteine may be considered as the 21st
amino acid.
4. Deficiency states:

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a. In Keshan province in China, the soil is deficient in selenium. This leads to
prevalence of Keshan disease. It is characterized by multifocal myocardial
necrosis, cardiac arrhythmias and cardiac enlargement. Selenium is known
to cure the disease.
b. Isolated selenium deficiency in other parts of the world caused liver
necrosis, cirrhosis, cardiomyopathy and muscular dystrophy. Selenium
toxicity is called selenosis.
5. Toxicity:
a. Selenium is present in metal polishes and anti-rust compounds. The toxicity
symptoms include hair loss, falling of nails, diarrhea, weight loss, and
garlicky odor in breath. The last mentioned symptom is due to the presence
of dimethyl selenide in expired air.
b. Kaschinbeck disease is characterized by degenerative osteoarthrosis.

1.
2.
3.
4.
5.

Nutrition,
Instrumentation & techniques used in biochemistry,
Thyroid, adrenocortical and renal function tests,
Interpretation of laboratory data,
Xenobiotics,

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6. Environmental hazards,
7. Cancer biology
Section II
1. Metabolism of proteins, amino acids,
2. Hemoglobin
3. Nucleotides, purines and pyrimidines, their regulation and associated inborn
errors
4. Porphyrias
5. Human genetics
6. Molecular biology
7. immunology