Purification and NH2-terminal amino acid sequencing of the β subunit of a human T-cell antigen receptor

O. Acuto, M. Fabbi, J. Smart

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75 Citations (Scopus)

Abstract

To obtain information about the structural basis for T-cell antigen recognition, a T3-associated Ti receptor molecule was isolated from crude membranes of the REX human thymic tumor line and purified by affinity chromatography with an anti-clonotypic monoclonal antibody in conjunction with preparative gel electrophoresis. NH2-terminal amino acid sequencing of the β subunit unambiguously identified the amino acids in positions 2-12. Comparative protein sequence analysis by computer search demonstrated that this Ti β sequence bore weak, but definite, homology to the first framework of the variable region of human λ light chain. Antisera to a synthetic peptide corresponding to positions 2-11 precipitated the denatured Ti β subunit from REX, thus confirming the above sequence. This information suggests that the Ti β subunit is distantly related to human immunoglobulin λ light chain and, moreover, should be of use in the molecular cloning of the Ti β gene.

Original languageEnglish
Pages (from-to)3851-3855
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number12 I
Publication statusPublished - 1984

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Protein Sequence Analysis
T-Cell Antigen Receptor
Thymus Neoplasms
Immunoglobulin Light Chains
Viral Tumor Antigens
Molecular Cloning
Affinity Chromatography
Electrophoresis
Immune Sera
Gels
Monoclonal Antibodies
T-Lymphocytes
Light
Amino Acids
Peptides
Membranes
Genes

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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T1 - Purification and NH2-terminal amino acid sequencing of the β subunit of a human T-cell antigen receptor

AU - Acuto, O.

AU - Fabbi, M.

AU - Smart, J.

PY - 1984

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AB - To obtain information about the structural basis for T-cell antigen recognition, a T3-associated Ti receptor molecule was isolated from crude membranes of the REX human thymic tumor line and purified by affinity chromatography with an anti-clonotypic monoclonal antibody in conjunction with preparative gel electrophoresis. NH2-terminal amino acid sequencing of the β subunit unambiguously identified the amino acids in positions 2-12. Comparative protein sequence analysis by computer search demonstrated that this Ti β sequence bore weak, but definite, homology to the first framework of the variable region of human λ light chain. Antisera to a synthetic peptide corresponding to positions 2-11 precipitated the denatured Ti β subunit from REX, thus confirming the above sequence. This information suggests that the Ti β subunit is distantly related to human immunoglobulin λ light chain and, moreover, should be of use in the molecular cloning of the Ti β gene.

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