Dehomocysteinylation is catalysed by the sirtuin-2-like bacterial lysine deacetylase CobB

FEBS J. 2016 Nov;283(22):4149-4162. doi: 10.1111/febs.13912. Epub 2016 Nov 2.

Abstract

Hyperhomocysteinemia, which is characterized by elevated blood levels of the non-protein amino acid homocysteine (Hcy), is an independent risk factor for many diseases, including cardiovascular diseases, neurodegenerative diseases and birth defects. The incorporation of homocysteine into proteins, known as protein N-homocysteinylation, has been considered a major mechanism that contributes to hyperhomocysteinemia. However, the process of dehomocysteinylation, the N-homocysteinylation substrates and the regulatory enzyme(s) remain largely unknown. In this study, we observed that the dehomocysteinylation reaction is a spontaneous process that can be inhibited by blocking -SH groups, which have been demonstrated to be critical for non-enzymatic dehomocysteinylation reactions. We also report that CobB, a known Sir2-like bacterial lysine deacetylase, catalyzes lysine dehomocysteinylation reactions both in vitro and in vivo. Our work provides insight into how this non-enzymatic modification might be removed from affected proteins, supplies potential targets for developing identification methods for N-homocysteine proteins, and identifies CobB as the first prokaryotic dehomocysteinylation enzyme.

Keywords: CobB; N-homocysteinylation; dehomocysteinylation; sirtuin; sulfhydryl group.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Carboxylic Ester Hydrolases / genetics
  • Carboxylic Ester Hydrolases / metabolism*
  • Cattle
  • HEK293 Cells
  • Histone Deacetylases / metabolism*
  • Homocysteine / analogs & derivatives
  • Homocysteine / chemistry
  • Homocysteine / metabolism*
  • Humans
  • Immunoblotting
  • Kinetics
  • Lysine / metabolism
  • Mice
  • Models, Chemical
  • Molecular Structure
  • Mutation
  • NIH 3T3 Cells
  • Protein Processing, Post-Translational
  • Serum Albumin, Bovine / chemistry
  • Serum Albumin, Bovine / metabolism
  • Sirtuin 2 / metabolism*

Substances

  • Bacterial Proteins
  • Homocysteine
  • Serum Albumin, Bovine
  • homocysteine thiolactone
  • Carboxylic Ester Hydrolases
  • CobB protein, Salmonella enterica
  • SIRT2 protein, human
  • Sirtuin 2
  • Histone Deacetylases
  • Lysine