Preparation and crystallization of the disulfide-linked HLA-G dimer

Biochim Biophys Acta. 2006 May;1764(5):985-8. doi: 10.1016/j.bbapap.2005.10.006. Epub 2005 Oct 25.

Abstract

HLA-G is a non-classical MHC class I, which binds to inhibitory receptors, such as Leukocyte Ig-like receptors, to induce a wide range of tolerogenic immunological effects. HLA-G can be expressed as a disulfide-liked dimer both in solution and at the cell surface. However, the three-dimensional structure of the HLA-G dimer is unknown. Here, we report the crystallization of the disulfide-linked dimer form of HLA-G by adding dithiothreitol (DTT), enabling a 3.2-A data set to be collected. We also show that DTT promotes disulfide bond exchange of refolded HLA-G, whose free cysteine was protected, thus facilitating its dimerization. This technique could also be applied for disulfide-mediated dimer/multimer formation of refolded proteins harbouring free cysteines.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization
  • Dimerization
  • Disulfides / chemistry*
  • Dithiothreitol
  • HLA Antigens / chemistry*
  • HLA Antigens / isolation & purification*
  • HLA-G Antigens
  • Histocompatibility Antigens Class I / chemistry*
  • Histocompatibility Antigens Class I / isolation & purification*
  • Humans

Substances

  • Disulfides
  • HLA Antigens
  • HLA-G Antigens
  • Histocompatibility Antigens Class I
  • Dithiothreitol