Intracellular transport blockade caused by disruption of the disulfide bridge in the third external domain of major histocompatibility complex class I antigen.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 3 ·1986-02-00 ·Pages 757-61

Miyazaki J, Appella E, Ozato K

Abstract

The third external domain of major histocompatibility class I antigens has a highly conserved disulfide bridge between cysteine-203 and cysteine-259. To elucidate the functional significance of this disulfide bridge, we have produced a mutant H-2Ld gene by site-directed mutagenesis in which the codon for cysteine-203 is changed to a codon for serine, which is unable to form a disulfide bridge. The mutant H-2Ld gene was introduced into mouse L cells and its expression has been studied. No measurable expression of the H-2Ld antigen was detected on the cell surface of the transformants by antibody-binding assays. However, a large quantity of the mutant H-2Ld antigen was found in the cytoplasm of the transformants as observed by immunoprecipitation of metabolically labeled lysate and by immunocytochemistry of membrane-permeabilized cells, using an antibody specific for the first external domain of the H-2Ld antigen. The mutant antigen was glycosylated and associated, at least in part, with beta 2-microglobulin. Subcellular fractionation experiments indicated that the transport of the antigen was blocked between the endoplasmic reticulum and the plasma membrane. It is concluded that structural integrity of the third external domain is a prerequisite for intracellular transport of class I antigens. On the basis of these findings we suggest that the domain structure containing the disulfide bridge serves as a signal structure necessary for receptor-mediated intracellular transport and that this requirement is the evolutionary basis for high conservation of similar structures present throughout the immunoglobulin supergene family.

MeSH Terms

Animals Biological Transport Cell Membrane/metabolism Disulfides/metabolism Endoplasmic Reticulum/metabolism Escherichia coli/genetics Genes, MHC Class II H-2 Antigens/genetics,metabolism Humans Major Histocompatibility Complex Mice Mutation

Chemicals

Disulfides H-2 Antigens

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Miyazaki J

Appella E

Ozato K

References (41)

41 references, click to expand

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Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.83.3.757

Published

1986-02-00

Pages

757-61

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC322944

Subset

External Links

PubMed source DOI: 10.1073/pnas.83.3.757 PMC Full Text View journal details

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Intracellular transport blockade caused by disruption of the disulfide bridge in the third external domain of major histocompatibility complex class I antigen.

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