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

Mitsunori Shiroishi, Daisuke Kohda, Katsumi Maenaka

Research output: Contribution to journalArticle

7 Citations (Scopus)

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-Å 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.

Original languageEnglish
Pages (from-to)985-988
Number of pages4
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1764
Issue number5
DOIs
Publication statusPublished - May 1 2006

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HLA-G Antigens
Crystallization
Disulfides
Dimers
Dithiothreitol
Cysteine
Protein Refolding
Dimerization
Leukocytes
Proteins

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Genetics

Cite this

Preparation and crystallization of the disulfide-linked HLA-G dimer. / Shiroishi, Mitsunori; Kohda, Daisuke; Maenaka, Katsumi.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1764, No. 5, 01.05.2006, p. 985-988.

Research output: Contribution to journalArticle

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