T cell antigen receptor (TCR) transmembrane peptides colocalize with TCR, not lipid rafts, in surface membranes.
We have previously shown that a synthetic peptide termed core peptide (CP), which corresponds to a sequence within the transmembrane domain of the alpha chain of the T cell antigen receptor (TCR), can inhibit IL-2 production in antigen-stimulated T cells and can suppress inflammation in several T cell-mediated animal models of disease. As the first step in determining the mechanism of CP action, we examined the association of CP with the plasma membrane of human T cells using confocal microscopy. A homogeneous distribution of CP was observed in the plasma membrane of human T cells. This membrane localization was dependent on the presence of positive charges in the CP sequence. CP analogs, containing either neutral or negatively charged amino acids in place of the positive amino acid charges, did not localize within TCR membranes. Following antibody-induced TCR clustering, there was specific colocalization of CP with surface TCR. No association was observed with other cell surface receptors when similarly clustered. Since TCR activation leads to an increased movement of the receptor complex to cholesterol/glycosphingolipid (GSL) plasma membrane microdomains (rafts) we examined whether the association of CP with TCR was raft-driven. TCR clustering led only to a partial colocalization of TCRs with raft GSL, ganglioside GM1, and a complete colocalization of CP with TCRs. We conclude that CP associates specifically with plasma membrane TCRs and not raft lipids.
Department of Rheumatology, Westmead Hospital, Sydney, NSW 2145, Australia.
Xin M
XM
Wang
Julianne T
JT
Djordjevic
Veronica
V
Bender
Nicholas
N
Manolios
D000595:Amino Acid Sequence; D000818:Animals; D019921:Biotinylation; D002462:Cell Membrane / Q000737:chemistry*; D002478:Cells, Cultured; D006801:Humans; D006825:Hybridomas; D021962:Membrane Microdomains / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D018613:Microscopy, Confocal; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry*; D014407:Tumor Cells, Cultured
D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; C464994:core protein, human; C464998:core protein, mouse