Question

“Protein A” is phosphorylated at one amino acid by a kinase, “protein B”, and dephosphorylated by a phosphatase, “protein C”. You have purified all three of these proteins and want to determine the structural changes that occur when protein A is phosphorylated or dephosphorylated. Explain in detail how you would go about designing and conducting this investigation.

Answer 1

Solution: I will make use of electrophoretic separation for rsolving the problem.

Initially we can divide the experiment into 3 groups.

Group 1: In this group Protein A was kept alone.

Group 2: Protein A was treated with Protein B

Group 3 : Protein A was treated with Protein B. This was followed by tratment with Proteic C.

Protein A from all these three groups after treatment will be separated and subjected to gel electrophoresis with dinuclear bispyridylmethylamine–tyrosine–acrylamide complexes. The addition of dinuclear bispyridylmethylamine–tyrosine–acrylamide clearly enhances the mobility shift of phosphorylated protein A in comparison to the dephosphorylated protein A.