A Recombinant, Soluble, Single-Chain Class I Major Histocompatibility Complex Molecule with Biological Activity

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 89; no. 22; pp. 10658 - 10662
• Main Authors: Mage, Michael G., Lee, Li, Ribaudo, Randall K., Corr, Maripat, Kozlowski, Steven, McHugh, Louise, Margulies, David H.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.11.1992; National Acad Sciences; National Academy of Sciences
• Subjects: Animals; Antibodies; Antibodies, Monoclonal; Antigens; Base Sequence; beta 2-Microglobulin - genetics; beta 2-Microglobulin - immunology; Biological and medical sciences; Biology; Cell Division; Cell Line; Cell lines; DNA - genetics; DNA - isolation & purification; Enzyme-Linked Immunosorbent Assay; Epitopes; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Genes; Genes, MHC Class I; H-2 Antigens - genetics; H-2 Antigens - immunology; Histocompatibility antigens (hla, h-2 and other systems); L cells; Liver - immunology; Macromolecular Substances; Major histocompatibility complex genes; Mice; Mice, Inbred C57BL; Molecular chains; Molecular immunology; Molecular Sequence Data; Molecules; Oligodeoxyribonucleotides; Oligonucleotides, Antisense; Polymerase Chain Reaction; Protein Folding; Proteins; Recombinant Proteins - immunology; Restriction Mapping; T lymphocytes; Transfection; Antigen presentation; Peptides; Major histocompatibility system; Rodentia; H-2-System; Biological activity; Mechanism; Vertebrata; Mammalia; Mouse; Molecular association; Molecular complex; Recombinant protein; Class I histocompatibility antigen; β2-Microglobulin
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.89.22.10658

Heterodimeric class I major histocompatibility complex molecules, which consist of a 45-kDa heavy-chain and a 12-kDa β2-microglobulin (β2m) 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 β2m 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 β2m, implying that a heavy chain/peptide complex is not an obligate intermediate in the assembly of the heavy-chain/β2m/peptide heterotrimer. Single-chain major histocompatibility complex molecules uniformly loaded with peptide have potential uses for structural studies, toxin or fluor conjugates, and vaccines