Biochemical identification of a direct physical interaction between the CD4

p56lck and Ti(TcR)/CD3 complexes

Kristine E. Burgess, Andreani D. Odysseos, Craig Zalvan, Brian Druker, Paul Anderson, Stuart F. Schlossman, Christopher E. Rudd

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

The CD4 and CD8 antigens function in synergy with the TcR/CD3 complex in the generation of intracellular signals leading to T cell proliferation. The association of the protein-tyrosine kinase p56lck with CD4 and CD8 provides a potential mechanism in the generation of intracellular signals. Several studies have shown that CD4 can co-modulate with TcR/CD3 suggesting that these receptor complexes may associate on the surface of the T cell. Nevertheless, it has proven difficult to formally demonstrate a direct physical interaction between the CD4 and TcR/CD3 complexes using biochemical techniques. In this study, we have used the sensitivity of the in vitro kinase assay to show a direct physical linkage between the CD4 : p56lck complex and various CD3 subunits. Immunoprecipitation of CD4 from cell lysates derived from the T lymphoblastoid line HPB-ALL results in the co-purification of p56lck with an additional polypeptide at 20 kDa. Re-precipitation analysis and isoelectric focusing demonstrated that this band corresponds to the CD3 ε chain. An alternative approach which involves the labeling of microsomal membranes with [γ-32P]ATP revealed the presence of CD3 ε and ζ chains in anti-CD4 immunoprecipitates. By contrast, we were unable to demonstrate the association of the CD4 : p56lck and TcR/CD3 complex in resting peripheral blood lymphocytes. These data indicate that the CD4 : p56lck and TcR/CD3 complexes have the ability to form stable complexes on the surface of certain T cell lines.

Original languageEnglish (US)
Pages (from-to)1663-1668
Number of pages6
JournalEuropean Journal of Immunology
Volume21
Issue number7
StatePublished - Jul 1991
Externally publishedYes

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CD3 Antigens
T-Lymphocytes
CD8 Antigens
CD4 Antigens
Isoelectric Focusing
Immunoprecipitation
Protein-Tyrosine Kinases
Phosphotransferases
Adenosine Triphosphate
Cell Proliferation
Lymphocytes
Cell Line
Peptides
Membranes

ASJC Scopus subject areas

  • Immunology

Cite this

Burgess, K. E., Odysseos, A. D., Zalvan, C., Druker, B., Anderson, P., Schlossman, S. F., & Rudd, C. E. (1991). Biochemical identification of a direct physical interaction between the CD4: p56lck and Ti(TcR)/CD3 complexes. European Journal of Immunology, 21(7), 1663-1668.

Biochemical identification of a direct physical interaction between the CD4 : p56lck and Ti(TcR)/CD3 complexes. / Burgess, Kristine E.; Odysseos, Andreani D.; Zalvan, Craig; Druker, Brian; Anderson, Paul; Schlossman, Stuart F.; Rudd, Christopher E.

In: European Journal of Immunology, Vol. 21, No. 7, 07.1991, p. 1663-1668.

Research output: Contribution to journalArticle

Burgess, KE, Odysseos, AD, Zalvan, C, Druker, B, Anderson, P, Schlossman, SF & Rudd, CE 1991, 'Biochemical identification of a direct physical interaction between the CD4: p56lck and Ti(TcR)/CD3 complexes', European Journal of Immunology, vol. 21, no. 7, pp. 1663-1668.
Burgess, Kristine E. ; Odysseos, Andreani D. ; Zalvan, Craig ; Druker, Brian ; Anderson, Paul ; Schlossman, Stuart F. ; Rudd, Christopher E. / Biochemical identification of a direct physical interaction between the CD4 : p56lck and Ti(TcR)/CD3 complexes. In: European Journal of Immunology. 1991 ; Vol. 21, No. 7. pp. 1663-1668.
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