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• W1982441198 abstract "In T cells, the lipid raft-associated Lck is strongly tyrosine phosphorylated and has reduced enzymic activity in contrast with the detergent-soluble pool, which has substantial activity. Lck tagged at the C-terminus (Lck/V5-His) was efficiently captured by epitope-specific reagents from the detergent-soluble fraction but not from lipid rafts. Binding was restored following urea denaturation, suggesting that Lck/V5-His is in a ‘closed’ conformation in these domains. In agreement with this hypothesis, the Tyr505 → Phe/V5-His and Arg154 → Lys/V5-His mutants, which disrupt the SH2-Tyr505 intramolecular interaction, were efficiently precipitated from lipid rafts. In contrast to Lck, Fyn/V5-His was precipitated equally well from both fractions. In the LAT (linker of activated T cells)-deficient J.CaM2 cells, Tyr505 phosphorylation of raft-associated Lck was reduced whereas its enzymic activity was elevated. This correlated with decreased levels of raft-localized Csk (C-terminal Src kinase) kinase. Increased tyrosine phosphorylation of Lck was restored in LAT-reconstituted J.CaM2 cells suggesting that LAT negatively regulates Lck activity in lipid rafts. Co-immunoprecipitation experiments from Tyr505 → Phe/V5-His-expressing cells revealed that LAT preferentially interacts with the ‘open’ form of Lck in T cell raft domains. These results demonstrate that, unlike the non-raft pool, Lck in lipid rafts has a ‘closed’-inactive structure, and that LAT plays a role in maintaining this conformation, possibly by facilitating critical associations within lipid rafts via its capacity to interact with the ‘open’ form of the kinase." @default.
• W1982441198 created "2016-06-24" @default.
• W1982441198 creator A5059522525 @default.
• W1982441198 date "2003-05-01" @default.
• W1982441198 modified "2023-09-27" @default.
• W1982441198 title "Selective interaction of LAT (linker of activated T cells) with the open-active form of Lck in lipid rafts reveals a new mechanism for the regulation of Lck in T cells" @default.
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• W1982441198 doi "https://doi.org/10.1042/bj20021578" @default.
• W1982441198 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/1223349" @default.
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