AIIMS PG NOVEMBER 2004: SUB JECT: BIOCHEMISTRY Q 1 .

The normal vale of P50 on the oxy haemoglobin dissociation curve in an adult is a) 1.8 kPa b) 2.7 c) 3.6 d) 4.5

Q 2 .Which of the following amino acid is most likely to be found in the trans membrane region of a protein a) Lysine b) Arginine c) Leucine d) Aspartate

Q 3 . Which of the following statement about HbS is not true : a) HbS differs from HbA by the substitution of Valine for Glutamate in position 6 of Beta chain b) One altered peptide HbS migrate faster toward the threshold (-) than the corresponding peptide of HbS c) Binding of HbS to the deoxygenated HbS can extend the polymer nd cause sickling of RBC d) Lowering the concentration of deoxygenated HbS can prevent sickling Q 4 .During the dark phase of visual cycle ,which form of Vitamn-A combine with Opsin to make Rhodopsin . a) All trans retinal dehyde b) All trans retinol c) 11-Cis retinal dehyde

d) 11- Cis retinol Q 5 .The molecular weight of protein can be determined by a) Native PAGE b) Sodium dodecyl sulphate h-PAGE c) Isoelectric focussing d) Ion exchange chromatography

Q 6 .Ladder pattern of DNA electrophoresis in apoptosis is caused by the action of following enzyme: a) Endonuclease b) Trans glutaminase c) DNAse d) Caspase

ANSWERS AND EXPLANATIONS 1. c, Explanation: In the haemoglobin-oxygen dissociation curve, the haemoglobin tetramer can bind up to 4 molecules of oxygen in the iron containing sites of the heme molecule. When acid is produced in the tissue, the dissociation curve shifts to the right. Alkalosis has the opposite effect, reducing oxygen delivery. So, P50= 26mm Hg= 3.5 kPa. 2. c, Explanation: The transmembrane region of a protein includes most of the non-polar amino acids. Hence the answer to the question is leucine which is the most non-polar of the choice given. 3. c, Explanation: In HbS, the nonpolar amino acid valine is replaced by Glutamine of the beta subunit, generating a hydrophobic sticky patch on the surface of the beta subunit of both oxy-HbS and deoxy-HbS. Both HbA and HbS have a complementary sticky patch on their surfaces that is exposed only if they are deoxygenated. That is, at low PO2, deoxy HbS is polymerized to form long, insoluble fibres.

 So, binding of deoxy-HbA terminates fibre polymerization, so HbA lacks the second sticky patch necessary to bind another Hb molecule. These twisted helicle fibres distort the erythrocyte into a characteristic sickle shape, so it becomes vulnerable to spleen sinusoids. A low PO2 such as high altitude accelerates the tendency to polymerize. 4. c, Explanation: When vitamin A is taken, it is in all trans retinol form. It goes to pigment epithelium from systemic circulation. There it is isomerized spontaneously to form 11 cis retinol and oxidized to 11 cis retinal. This end product is combined with opsin to form rhodopsin which is deposited in the cell membrane of pigment epithelium of retina. Due to this holoenzyme formation, the chemical equilibrium of [All trans ----- 11 cis ] always tilt to the right to facilitate 11 cis conformation formation. When photon excites Rhodopsin, 11 cis retinal is converted to all trans form and for this retinal is released from apoprotein initiating a nerve impulse. Then a cascade of reaction forming rhodopsin, bathorhodopsin occurs in pico seconds. It results in ultimate formation of metarhodopsin 2 that causes cGMP formation as second messenger and initiate a nerve impulse via a intracellular signal transducing pathway. And the final step is hydrolysis of All trans retinal. So the key form of vitamin-A which combines with opsin, to initiate the Walds-Visual cycle, is 11 cis retinal.

5. B, SDS PAGE: Sodium dodesyl sulfate polyaccrilamide gel electrophoresis. Explanation: Here SDS is amphipathic in nature with negative charge at one end due to Sulfate Group. So it coats the polar peptides in an approximate proportion of one SDS per Two peptide bonds. This imparts huge negative charge on the protein irrespective of its amino acid sequence. Also due to huge amount of SDS combined with a specific protein, the mobility of electrophoresis depends on the number of SDS bound with a single protein which is also determined by the number of amino acid of the protein, a rough determinant of the molecular weight of the protein. Special note: If beta marcapto ethanol/ guanidium chloride is given along with SDS PAGE to disrupt the disulfide bonds and tertiary structures, the molecular weight of the peptides instead of the total protein is determined.

6. a, Explanation: The biochemical hallmark of apoptosis is fragmentation of the genomic DNA, an irreversible event that commits the cell to death. This fragmentation of DNA is achieved by Endoclease, which causes endogenous activation of Ca And Mg channels. Special note: The enzyme endonuclease cleaves the DNA between two linker DNA segments generating mono and di oligonucleosomal units.