Effect of leucine-to-methionine substitutions on the diffraction quality of histone chaperone SET/TAF-Ibeta/INHAT crystals

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Oct 1;64(Pt 10):960-5. doi: 10.1107/S1744309108028704. Epub 2008 Sep 30.

Abstract

One of the most frequent problems in crystallization is poor quality of the crystals. In order to overcome this obstacle several methods have been utilized, including amino-acid substitutions of the target protein. Here, an example is presented of crystal-quality improvement by leucine-to-methionine substitutions. A variant protein with three amino-acid substitutions enabled improvement of the crystal quality of the histone chaperone SET/TAF-Ibeta/INHAT when combined with optimization of the cryoconditions. This procedure improved the resolution of the SET/TAF-Ibeta/INHAT crystals from around 5.5 to 2.3 A without changing the crystallization conditions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Chromosomal Proteins, Non-Histone / chemistry
  • Chromosomal Proteins, Non-Histone / genetics*
  • Chromosomal Proteins, Non-Histone / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • DNA-Binding Proteins
  • Genetic Variation
  • Histone Chaperones
  • Histones / metabolism
  • Humans
  • Leucine / metabolism*
  • Methionine / metabolism*
  • Molecular Chaperones / chemistry
  • Molecular Chaperones / genetics*
  • Molecular Chaperones / metabolism
  • Sensitivity and Specificity
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism

Substances

  • Chromosomal Proteins, Non-Histone
  • DNA-Binding Proteins
  • Histone Chaperones
  • Histones
  • Molecular Chaperones
  • SET protein, human
  • Transcription Factors
  • Methionine
  • Leucine