A recombinant, soluble, single-chain class I major histocompatibility complex molecule with biological activity.
Heterodimeric class I major histocompatibility complex molecules, which consist of a 45-kDa heavy-chain and a 12-kDa beta 2-microglobulin (beta 2m) light chain, bind endogenously synthesized peptides for presentation to antigen-specific T cells. We have synthesized a gene encoding a single-chain, soluble class I molecule derived from mouse H-2Dd, in which the carboxyl terminus of beta 2m is linked via a peptide spacer to the amino terminus of the heavy chain. The chimeric protein is secreted efficiently from transfected L cells, is thermostable, and when loaded with an appropriate antigenic peptide, stimulates an H-2Dd-restricted antigen-specific T-cell hybridoma. Thus, functional binding of peptide does not require the complete dissociation of beta 2m, implying that a heavy chain/peptide complex is not an obligate intermediate in the assembly of the heavy-chain/beta 2m/peptide heterotrimer. Single-chain major histocompatibility complex molecules uniformly loaded with peptide have potential uses for structural studies, toxin or fluor conjugates, and vaccines.
Mage
M G
MG
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Lee
L
L
Ribaudo
R K
RK
Corr
M
M
Kozlowski
S
S
McHugh
L
L
Margulies
D H
DH
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002455:Cell Division; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D005805:Genes, MHC Class I; D006183:H-2 Antigens / Q000235:genetics / Q000276:immunology*; D008099:Liver / Q000276:immunology; D046911:Macromolecular Substances; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D016133:Polymerase Chain Reaction; D017510:Protein Folding; D011994:Recombinant Proteins / Q000276:immunology; D015183:Restriction Mapping; D014162:Transfection; D001613:beta 2-Microglobulin / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006183:H-2 Antigens; D046911:Macromolecular Substances; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D001613:beta 2-Microglobulin; D004247:DNA