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PMID: 8978819 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Exclusion of CD45 inhibits activity of p56lck associated with glycolipid-enriched membrane domains.

The Journal of cell biology ·Vol. 135 ·No. 6 Pt 1 ·1996-12-00 ·Pages 1515-23

Rodgers W, Rose JK

Abstract

p56lck (Lck) is a lymphoid-specific Src family tyrosine kinase that is critical for T-cell development and activation. Lck is also a membrane protein, and approximately half of the membrane-associated Lck is associated with a glycolipid-enriched membrane (GEM) fraction that is resistant to solubilization by Triton X-100 (TX-100). To compare the membrane-associated Lck present in the GEM and TX-100-soluble fractions of Jurkat cells, Lck from each fraction was immunoblotted with antibody to phosphotyrosine. Lck in the GEM fraction was found to be hyperphosphorylated on tyrosine, and this correlated with a lower kinase specific activity relative to the TX-100-soluble Lck. Peptide mapping and phosphatase diagests showed that the hyperphosphorylation and lower kinase activity of GEM-associated Lck was due to phosphorylation of the regulatory COOH-terminal Tyr505. In addition, we determined that the membrane-bound tyrosine phosphatase CD45 was absent from the GEM fraction. Cells lacking CD45 showed identical phosphorylation of Lck in GEM and TX-100-soluble membranes. We propose that the GEM fraction represents a specific membrane domain present in T-cells, and that the hyperphosphorylation of tyrosine and lower kinase activity of GEM-associated Lck is due to exclusion of CD45 from these domains. Lck associated with the GEM domains may therefore consitute a reservoir of enzyme that can be readily activated.

MeSH Terms
Cell Fractionation Cell Membrane/metabolism Glycolipids/metabolism HeLa Cells Humans Jurkat Cells Leukocyte Common Antigens/metabolism Lymphocyte Specific Protein Tyrosine Kinase p56(lck) Membrane Lipids/metabolism Phosphorylation Tyrosine/metabolism src-Family Kinases/antagonists & inhibitors,metabolism
Chemicals
Glycolipids Membrane Lipids Tyrosine Lymphocyte Specific Protein Tyrosine Kinase p56(lck) src-Family Kinases Leukocyte Common Antigens
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Rodgers W
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.
Rose J K
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.6.1515
Published
1996-12-00
Pages
1515-23
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2133949
Subset
IM
Grants
NIAID NIH HHS · AI 24345 · United States
External Links
PubMed source DOI: 10.1083/jcb.135.6.1515 PMC Full Text View journal details
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