Direct determination of the redox status of cysteine residues in proteins in vivo

Biochem Biophys Res Commun. 2015 Jan 2;456(1):339-43. doi: 10.1016/j.bbrc.2014.11.082. Epub 2014 Nov 28.

Abstract

The redox states of proteins in cells are key factors in many cellular processes. To determine the redox status of cysteinyl thiol groups in proteins in vivo, we developed a new maleimide reagent, a photocleavable maleimide-conjugated single stranded DNA (DNA-PCMal). The DNA moiety of DNA-PCMal is easily removed by UV-irradiation, allowing DNA-PCMal to be used in Western blotting applications. Thereby the state of thiol groups in intracellular proteins can be directly evaluated. This new maleimide compound can provide information concerning redox proteins in vivo, which is important for our understanding of redox networks in the cell.

Keywords: Cysteine; Photocleavable maleimide-conjugated DNA; Sulfhydryl; Thiol reagent; Thiol redox status; Western blotting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology
  • Blotting, Western
  • Cysteine / chemistry*
  • DNA / chemistry*
  • Electrophoresis, Gel, Two-Dimensional
  • Escherichia coli / enzymology
  • HeLa Cells
  • Humans
  • Maleimides / chemistry*
  • Oxidation-Reduction*
  • Oxidative Stress
  • Proteins / chemistry*
  • Sulfhydryl Compounds / chemistry*
  • Ultraviolet Rays

Substances

  • Maleimides
  • Proteins
  • Sulfhydryl Compounds
  • maleimide
  • DNA
  • Cysteine