title: Signal transduction and glycophosphatidylinositol-linked proteins (lyn, lck, CD4, CD45, G proteins, and CD55) selectively localize in Triton-insoluble plasma membrane domains of human leukemic cell lines and normal granulocytes.
abstract: Src-family nonreceptor protein tyrosine kinases (NRPTK) are associated with cell surface receptors in large detergent-resistant complexes: in epithelial cells, yes is selectively located in vesicle structures containing caveolin ("caveolae"). These formations are typically also endowed with glycophosphatidylinositol (GPI)-anchored proteins. In the present study, we observed lck, lyn, src, hck, CD4, CD45, G proteins, and CD55 (decay-accelerating factor) expression in the buoyant low-density Triton-insoluble (LDTI) fraction of selected leukemic cell lines and granulocytes. We provide a detailed analysis of the two most highly expressed NRPTK, p53/p56lyn and p56lck, which are involved in the transduction of signals for proliferation and differentiation of monocytes/B lymphocytes and T lymphocytes, respectively. We show that lyn is selectively recovered in LDTI complexes isolated from human leukemic cell lines (promyelocytic [HL-60], erythroid [K562] and B-lymphoid [697]) and from normal human granulocytes, and that lck is recovered from LDTI fractions of leukemic T- and B-lymphoid cell lines (CEM, 697). In LDTI fractions of leukemic cells, lck and lyn are enriched 100-fold as compared with the total cell lysates. Analysis of these fractions by electron microscopy shows the presence of 70- to 200-nm vesicles: lyn and lck are homogenously distributed in the vesicles, as revealed by an immunogold labeling procedure. These novel results propose a role for these vesicles in signal transduction mechanisms of normal and neoplastic hematopoietic cells. In support of this hypothesis, we further observed that molecules participating in B- and T-cell receptor activation cofractionate in the LDTI fractions, CD45/lyn (B cells) and CD45/lck/CD4 (T cells).
authors.affiliation: Thomas Jefferson Cancer Institute, Thomas Jefferson University, Philadelphia, PA 19107-5541, USA.
authors.forename: I
authors.initials: I
authors.lastname: Parolini
authors.forename: M
authors.initials: M
authors.lastname: Sargiacomo
authors.forename: M P
authors.initials: MP
authors.lastname: Lisanti
authors.forename: C
authors.initials: C
authors.lastname: Peschle
mesh_terms: D015703:Antigens, CD / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006098:Granulocytes / Q000378:metabolism*; D006801:Humans; D007938:Leukemia / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D017830:Octoxynol; D015398:Signal Transduction
chemical_list: D015703:Antigens, CD; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D017830:Octoxynol