Saturation mutagenesis of a major histocompatibility complex protein domain: Identification of a single conserved amino acid important for allorecognition

R. Murray, C. A. Hutchison, J. A. Frelinger

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The interactive association between T lymphocytes and their target cells is an important system of cell-cell interactions. Major histocompatibility complex class I molecules are the cell surface structures recognized by cytolytic T lymphocytes. To define the molecular structures recognized by cytotoxic T lymphocytes, we have saturated the 270-base-pair α1 exon of the H-2D(p) gene with point mutations, rapidly producing a 'library' of 2.5 x 103 independent mutants. The library contains enough recombinant clones (each clone encoding approximately one amino acid replacement mutation) to predict a mutation at each nucleotide position of the α1 exon. The functional analysis of the first five transfected gene products tested has shown that mutation of a conserved tyrosine at position 27 to asparagine destroys recognition of the H-2D(p) gene product by polyclonal alloreactive cytotoxic T lymphocytes. Recognition of the same mutant molecule by three monoclonal antibodies and H-2-restricted lymphocytic choriomeningitis virus-specific cytotoxic T lymphocytes is unaffected.

Original languageEnglish (US)
Pages (from-to)3535-3539
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume85
Issue number10
DOIs
StatePublished - 1988
Externally publishedYes

ASJC Scopus subject areas

  • General

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