Detection of phosphorylation by enzymatic techniques.

Published

Journal Article

Reversible protein phosphorylation is an important mechanism for regulating physiological processes in both plant and animal cells. There are a number of techniques to demonstrate the presence of covalently bound phosphate in proteins. The general strategy of the protocols in this unit is to first examine the functional effects elicited by nonspecific acid or alkaline phosphatases that dephosphorylate many phosphoproteins in vitro. Protein phosphatases that selectively hydrolyze phosphoserine and phosphothreonine or phosphotyrosine residues can then be used to identify a functionally important covalent modification. Additional protocols describe digestion of phosphoproteins with a protein serine/threonine phosphatase and protein tyrosine phosphatase. A support protocol has been included to identify the radiolabel as (32)Pi based on its ability to form a complex with ammonium molybdate.

Full Text

Duke Authors

Cited Authors

  • Shenolikar, S

Published Date

  • May 2001

Published In

Volume / Issue

  • Chapter 18 /

Start / End Page

  • Unit - 18.5

PubMed ID

  • 18265170

Pubmed Central ID

  • 18265170

Electronic International Standard Serial Number (EISSN)

  • 1934-3647

Digital Object Identifier (DOI)

  • 10.1002/0471142727.mb1805s33

Language

  • eng

Conference Location

  • United States