Biochem review session 9 3 Possible test questions 1 What properties of proteins or peptides are derived from these procedures or determine the behavior a SDS gel b Isoelectric focusing separating molecules by differences in their isoelectric points c Gel chromatography d Ion exchange chromatography e Reverse phase high performance liquid chromatography RD HPLC 2 What is the effect of urea or guanidine HCl on proteins How was this important in the Antinsen s experiments on RNAse A a Abolish tertiary and secondary structures destroy them 3 Match 20 points a Edman degredation b RP HPLC c Gel chromatography d Ion exchange chromatography e PAGE polyacrylamide gel electrophoresis f SDS gel electrophoresis g Trypsin digestion i Criterion of purity c d e ii Size separation c f iii Subunit molecular weight f iv PITC a v Concentration vi Separation by net charge d vii Peptide separation b viii Dialysis 4 On gel filtration of the following proteins in what order will they probably elute from the column a RNAse A M 13 KD 3rd b Carbonic anhydrase M 29 kD 2nd c Bovine serum albumin M 66 kD 1st 5 On electrophoresis on an SDS gel after all disulfides are reduced in what order will these proteins all single chains move a Larger proteins move slower depends on molecular weight smaller protein moves fastest on gel sheet a moves fastest 6 Given the polypeptide ala phe glu lys thr gly cys trp ala pro met ala gly a Digest with i Trypsin ii Chymotrypsin iii V8 protease iv CNBr 1 Indicate the cleavage site with vertical arrows and the appropriate letter in pictures 7 Fill in the correct letters 20 points a Mb ii iv v b Hb i iii v vi i O2 carrier in blood ii O2 carrier in muscle iii Alpha2 beta2 subunit structure iv Single polypeptide chin v Heme prosthetic group vi Cooperative binding of O2 vii Allosteric control by H CO2 2 3BPG viii First x my structure ix M 18 kD