Chitin and Stress Induced Protein Kinase Activation

Methods Mol Biol. 2017:1578:185-194. doi: 10.1007/978-1-4939-6859-6_15.

Abstract

The assays described here are pertinent to protein kinase studies in any plant. They include an immunoblot phosphorylation/activation assay and an in-gel activity assay for MAP kinases (MPKs) using the general protein kinase substrate myelin basic protein. They also include a novel in-gel peptide substrate assay for Snf1-related kinase family 2 members (SnRK2s). This kinase family-specific assay overcomes some limitations of in-gel assays and permits the identification of different types of kinase activities in total protein extracts.

Keywords: Acryloylated peptide substrates; In-gel assays; Myelin basic protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chitin / pharmacology*
  • Enzyme Activation
  • Immunoblotting
  • Mitogen-Activated Protein Kinases / metabolism*
  • Myelin Basic Protein / metabolism*
  • Phosphorylation
  • Plant Proteins / metabolism
  • Plants / drug effects*
  • Plants / enzymology
  • Stress, Physiological
  • Substrate Specificity

Substances

  • Myelin Basic Protein
  • Plant Proteins
  • Chitin
  • Mitogen-Activated Protein Kinases