Structural requirements for the enzymatic phosphorylation of phosvitin.
Some structural features required for the enzymatic phosphorylation of phosvitin by purified rat liver cytosol phosvitin kinase have been investigated by testing the activity of such an enzyme toward phosphopeptides differing in size and chemical composition, obtained by pronase or acid hydrolysis of phosvitin. The results obtained can be summarized as follows: (a) Phosvitin kinase phosphorylates even fairly simple phosphopeptides (mol.wt 1000-2000) at rates comparable with intact phosvitin. (b) Acetylation of both phosvitin and pronase phosphopeptides completely prevents their phosphorylation indicating that some lysine residues are strictly required for the phosvitin kinase reaction. (c) Accordingly polyphosphorylserine blocks Ser(P)n which are very actively phosphorylated in phosvitin and pronase phosphopeptides, do not undergo any more enzymatic phosphorylation once isolated as such in a form free of other amino acids. (d) The activity of phosvitin kinase toward substrates probably devoid of Ser(P)n blocks suggests that there are not required for the protein kinase reaction. However, they apparently enhance the phosphorylation rate of the peptide substrates, likely by making easier their binding to the enzyme. It is proposed therefore that the peptidic unit able to undergo phosphorylation by rat liver cytosol phosvitin kinase consists of one or more phosphorylserine residues having in their close proximity a lysine residue playing a critical role in the mechanism of transphosphorylation.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004527:Egg Proteins; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010718:Phosphatidylserines / Q000494:pharmacology; D010774:Phosvitin; D011402:Pronase; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats
D000596:Amino Acids; D004527:Egg Proteins; D010446:Peptide Fragments; D010718:Phosphatidylserines; D010774:Phosvitin; D011494:Protein Kinases; D011402:Pronase