A comparative study of ATP analogs for phosphorylation-dependent kinase-substrate crosslinking

Bioorg Med Chem. 2014 Mar 1;22(5):1620-5. doi: 10.1016/j.bmc.2014.01.034. Epub 2014 Jan 30.

Abstract

Kinase-catalyzed protein phosphorylation is an important post-translational modification that regulates a variety of cellular functions. Identification of the many substrates of a specific kinase is critical to fully characterize cell biology. Unfortunately, kinase-substrate interactions are often transient, which makes their identification challenging. Here, the transient kinase-substrate complex was stabilized by covalent crosslinking using γ-phosphate modified ATP analogs. Building upon prior use of an ATP-aryl azide photocrosslinking analog, we report here the creation of an ATP-benzophenone photocrosslinking analog. ATP-benzophenone displayed a higher conversion percentage but more diffuse crosslinking compared to the ATP-aryl azide analog. A docking study was also performed to rationalize the conversion and crosslinking data. In total, the photocrosslinking ATP analogs produced stable kinase-substrate complexes that are suitable for future applications characterizing cell signaling pathways.

Keywords: ATP analogs; Kinase; Kinase substrates; Photocrosslinking.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analogs & derivatives*
  • Adenosine Triphosphate / metabolism
  • Humans
  • Phosphorylation / physiology*
  • Protein Kinases / metabolism*
  • Protein Processing, Post-Translational
  • Signal Transduction
  • Substrate Specificity

Substances

  • Adenosine Triphosphate
  • Protein Kinases