5. The protein allergen from peanuts, a protein called Ara h8 was recently purified and characterized. The investigators initially had difficulty separating Ara h8 from a similar protein in peanuts called Ara h6 because the two proteins were of similar size and had nearly identical pl values. Separation of the two proteins was finally achieved when it was noted that the Ara h6 protein contained ten cysteine residues involved in disulfide bridges, whereas Ara h8 contained no cysteines. The protein mixture was treated with a reducing agent, dithiothreitol (DTT), and then treated with iodoacetic acid (ICH₂COOH, a reagent that adds to, or alkylates, an-SH group and releases free iodine). The mixture was then loaded onto an anion exchange column and the two proteins were successfully separated. a) Show the structural changes that occur when Cys residues are exposed to DTT followed by iodoacetic acid.
5. The protein allergen from peanuts, a protein called Ara h8 was recently purified and characterized. The investigators initially had difficulty separating Ara h8 from a similar protein in peanuts called Ara h6 because the two proteins were of similar size and had nearly identical pl values. Separation of the two proteins was finally achieved when it was noted that the Ara h6 protein contained ten cysteine residues involved in disulfide bridges, whereas Ara h8 contained no cysteines. The protein mixture was treated with a reducing agent, dithiothreitol (DTT), and then treated with iodoacetic acid (ICH₂COOH, a reagent that adds to, or alkylates, an-SH group and releases free iodine). The mixture was then loaded onto an anion exchange column and the two proteins were successfully separated. a) Show the structural changes that occur when Cys residues are exposed to DTT followed by iodoacetic acid.
Basic Clinical Laboratory Techniques 6E
6th Edition
ISBN:9781133893943
Author:ESTRIDGE
Publisher:ESTRIDGE
Chapter6: Basic Clinical Chemistry
Section6.1: Introduction To Clinical Chemistry
Problem 9RQ
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