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 language | English |
|---|---|
| Pages (from-to) | 985-988 |
| Number of pages | 4 |
| Journal | Biochimica et Biophysica Acta - Proteins and Proteomics |
| Volume | 1764 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - May 1 2006 |
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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 journal › Article
}
TY - JOUR
T1 - Preparation and crystallization of the disulfide-linked HLA-G dimer
AU - Shiroishi, Mitsunori
AU - Kohda, Daisuke
AU - Maenaka, Katsumi
PY - 2006/5/1
Y1 - 2006/5/1
N2 - 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.
AB - 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.
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UR - http://www.scopus.com/inward/citedby.url?scp=33646510515&partnerID=8YFLogxK
U2 - 10.1016/j.bbapap.2005.10.006
DO - 10.1016/j.bbapap.2005.10.006
M3 - Article
C2 - 16290109
AN - SCOPUS:33646510515
VL - 1764
SP - 985
EP - 988
JO - Biochimica et Biophysica Acta - Proteins and Proteomics
JF - Biochimica et Biophysica Acta - Proteins and Proteomics
SN - 1570-9639
IS - 5
ER -