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Recent evidence suggests that peptide ligands take part in the assembly of class I molecules in living cells. We now describe a simple system for studying class I assembly in vitro. Detergent extracts of the mutant cells RMA-S and .174, in which class I assembly does not occur spontaneously, will support assembly in vitro when specific peptides are added. Peptides stabilize a conformational change in the class I heavy chain and association with beta 2-microglobulin, at concentrations approximately 100-fold lower than required in "peptide feeding" experiments with whole cells. We show that peptides bind class I molecules during assembly and demonstrate that the conformational change induced in the heavy chain is influenced by the concentrations of both peptide and beta 2-microglobulin.

Type

Journal article

Journal

Cell

Publication Date

27/07/1990

Volume

62

Pages

285 - 295

Keywords

Amino Acid Sequence, Animals, Cell-Free System, Histocompatibility Antigens Class I, Humans, In Vitro Techniques, Kinetics, Macromolecular Substances, Mice, Molecular Sequence Data, Peptides, Protein Conformation, beta 2-Microglobulin