I need help with number 6. protein hormone insulin stimulates uptake of glucose

Question

I need help with number 6.

protein hormone insulin stimulates uptake of glucose from biood into tissues. Deficiencies in insulin production or response result in the diease diabetes. Insulin consists of two peptide chains (refered to as the A chais asdt B chain) joined together by disulfide bonds. The following table shows sequence variation among various animal insulins Table 4.1: Varien Species B2 B27 0 A8 Thr Ser e Ala -human Ser Val Ser Ile Thr Gly lle Se Ilie Ser lle Asn Th Val Thr Ala Val The Ala Val Thr Ser pig rabbit Thr Phe dog -chicken Ala Ala Ser Ala Ala Ser Thr Glu Ala duck Which animal insulin would be the most compatible choice for treating a diabetic human? A) cow pig C) chickern D) duck 6. Asparagine and glutamine residues in proteins will sometimes be deamidated into aspartic acid and glutamic acid, respectively. If this happens to a purified protein, it will result in a mixed population of proteins with the original Asn and Gin resides and proteins in which they have been replaced by Asp and Glu. What would be the best technique to separate original wild-type proteins from the deamidated ones? SDS-PAGE Solubility assays (salting out) C) Gel filtration D) lon exchange

Explanation / Answer

Answer 6) In a mixture of protein with original Asn and Gln residue and protein with deamidated residues aspartic acid and Glutamic acid will have different charge on their surface because of difference in their amino acid sequences. This is because aspartic acid and Glutamic acid will contribute negative charge to the protein. Thus when they are mixed with salt in aqueous solution, both protein will precipitate at different concentration of salts. So we salting out would be the best technique to separate both proteins.

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