The site-specific installation of methyl-lysine analogs into recombinant histones

Cell. 2007 Mar 9;128(5):1003-12. doi: 10.1016/j.cell.2006.12.041.

Abstract

Histone lysine residues can be mono-, di-, or trimethylated. These posttranslational modifications regulate the affinity of effector proteins and may also impact chromatin structure independent of their role as adaptors. In order to study histone lysine methylation, particularly in the context of chromatin, we have developed a chemical approach to install analogs of methyl lysine into recombinant proteins. This approach allows for the rapid generation of large quantities of histones in which the site and degree of methylation can be specified. We demonstrate that these methyl-lysine analogs (MLAs) are functionally similar to their natural counterparts. These methylated histones were used to examine the influence of specific lysine methylation on the binding of effecter proteins and the rates of nucleosome remodeling. This simple method of introducing site-specific and degree-specific methylation into recombinant histones provides a powerful tool to investigate the biochemical mechanisms by which lysine methylation influences chromatin structure and function.

Publication types

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

MeSH terms

  • Animals
  • Cysteine / chemistry
  • Histones / chemistry*
  • Histones / genetics
  • Histones / metabolism
  • Lysine / analogs & derivatives*
  • Methylation
  • Models, Molecular
  • Nucleosomes / chemistry
  • Nucleosomes / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Xenopus Proteins / chemistry*
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism

Substances

  • Histones
  • Nucleosomes
  • Recombinant Proteins
  • Xenopus Proteins
  • Lysine
  • Cysteine