Translocation of peptides through microsomal membranes is a rapid process and promotes assembly of HLA-B27 heavy chain and beta 2-microglobulin translated in vitro.

Details

Serval ID
serval:BIB_A27FA717473B
Type
Article: article from journal or magazin.
Collection
Publications
Title
Translocation of peptides through microsomal membranes is a rapid process and promotes assembly of HLA-B27 heavy chain and beta 2-microglobulin translated in vitro.
Journal
The Journal of cell biology
Author(s)
Lévy F., Larsson R., Kvist S.
ISSN
0021-9525 (Print)
ISSN-L
0021-9525
Publication state
Published
Issued date
11/1991
Peer-reviewed
Oui
Volume
115
Number
4
Pages
959-970
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
We have translated major histocompatibility complex (MHC) class I heavy chains and human beta 2-microglobulin in vitro in the presence of microsomal membranes and a peptide from the nucleoprotein of influenza A. This peptide stimulates assembly of HLA-B27 heavy chain and beta 2-microglobulin about fivefold. By modifying this peptide to contain biotin at its amino terminus, we could precipitate HLA-B27 heavy chains with immobilized streptavidin, thereby directly demonstrating class I heavy chain-peptide association under close to physiological conditions. The biotin-modified peptide stimulates assembly to the same extent as the unmodified peptide. Both peptides bind to the same site on the HLA-B27 molecule. Immediately after synthesis of the HLA-B27 heavy chain has been completed, it assembles with beta 2-microglobulin and peptide. These interactions occur in the lumen of the microsomes (endoplasmic reticulum), demonstrating that the peptide must cross the microsomal membrane in order to promote assembly. The transfer of peptide across the microsomal membrane is a rapid process, as peptide binding to heavy chain-beta 2-microglobulin complexes is observed in less than 1 min after addition of peptide. By using microsomes deficient of beta 2-microglobulin (from Daudi cells), we find a strict requirement of beta 2-microglobulin for detection of peptide interaction with the MHC class I heavy chain. Furthermore, we show that heavy chain interaction with beta 2-microglobulin is likely to precede peptide binding. Biotin-modified peptides are likely to become a valuable tool in studying MHC antigen interaction and assembly.
Keywords
Amino Acid Sequence, Biological Transport, Biotin, Cell-Free System, Culture Techniques, Endoplasmic Reticulum/metabolism, HLA-B27 Antigen/biosynthesis, Histocompatibility Antigens Class I/biosynthesis, Humans, Intracellular Membranes/metabolism, Kinetics, Microsomes/metabolism, Molecular Sequence Data, Nucleoproteins/metabolism, Peptides/metabolism, Protein Biosynthesis, beta 2-Microglobulin/biosynthesis
Pubmed
Web of science
Open Access
Yes
Create date
28/01/2008 12:17
Last modification date
01/07/2024 17:36
Usage data