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Reconstitution of class I MHC molecules expressed in E. coli and complexed with single antigenic peptides

Authors
 Jongsun Kim  ;  Mi-Yeon Kim  ;  Yong Joon Chwae  ;  Jeon Han Park 
Citation
 JOURNAL OF KOREAN MEDICAL SCIENCE, Vol.12(4) : 280-285, 1997 
Journal Title
 JOURNAL OF KOREAN MEDICAL SCIENCE 
ISSN
 1011-8934 
Issue Date
1997
MeSH
Crystallization ; Escherichia coli ; HLA-A2 Antigen/isolation & purification ; Histocompatibility Antigens Class I/biosynthesis ; Histocompatibility Antigens Class I/genetics* ; Histocompatibility Antigens Class I/isolation & purification ; Humans ; Peptides/immunology* ; Recombinant Proteins/biosynthesis ; Recombinant Proteins/isolation & purification
Abstract
The HLA-Cw3 heavy chain has been expressed at high level as insoluble protein aggregates in E. coli. The protein aggregates dissolved in strong denaturant solution were efficiently reconstituted by removal of denaturant in the presence of an HLA-Cw3 binding peptide (FAM) and beta 2m. The reconstituted HLA-Cw3/FAM protein binds specifically to a p58 natural killer cell inhibitory receptor, a natural ligand. The HLA-A2 molecule has also been reconstituted in complex with either of a peptide from myelin associated glycoprotein (MAG) or a peptide from the GAG protein of human immunodeficiency virus. The HLA-A2/MAG protein crystallized under the identical conditions as HLA-A2 purified from human lymphoblastoid cells. The reconstitution method has yielded an abundant supply of HLA molecules complexed with single antigenic peptides, and may be of general utility in reconstituting any class I MHC molecules. However, the HLA molecules could not be reconstituted either without a peptide or with an irrelevant peptide. Using this property, the reconstitution method could be used to determine whether a peptide is restricted/bound to certain class I MHC molecule.
Files in This Item:
T199701523.pdf Download
DOI
10.3346/jkms.1997.12.4.280
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Microbiology (미생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Jong Sun(김종선) ORCID logo https://orcid.org/0000-0002-3149-669X
Park, Jeon Han(박전한) ORCID logo https://orcid.org/0000-0001-9604-3205
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/177333
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