Characterization of an associated 16-kDa tyrosine phosphoprotein required for Ly-49D signal transduction.
Ly-49D is an activating receptor on NK cells that does not become tyrosine phosphorylated upon activation. This report demonstrates that immunoprecipitation of Ly-49D, following pervanadate treatment or specific Ab cross-linking, coprecipitates a 16-kDa tyrosine-phosphorylated protein (pp16). Immunoblotting experiments and data from TCR-zeta/Fc epsilonRIgamma double knockout mice confirm that pp16 is not TCR-zeta, TCR-eta, or Fc epsilonRIgamma. Association of pp16 with Ly-49D involves a transmembrane arginine since mutation to leucine (Ly-49D[R54L]) abolishes association with pp16 in transfected P815 cells. In addition, Ly-49D(R54L) transfectants fail to mediate Ca2+ mobilization following Ab cross-linking. Therefore, signaling through Ly49D on NK cells depends on association with a distinct tyrosine phosphoprotein (pp16) in a manner analogous to that of TCR and FcR. Expression of this novel signaling peptide in both the NK and myeloid lineages indicates that pp16 is likely involved in the signal transduction cascade of additional receptor families.
Mason
L H
LH
Laboratory of Experimental Immunology, Division of Basic Sciences, SAIC Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, MD 21702, USA. MASON@MAIL.NCIFCRF.GOV
Willette-Brown
J
J
Anderson
S K
SK
Gosselin
P
P
Shores
E W
EW
Love
P E
PE
Ortaldo
J R
JR
McVicar
D W
DW
D000818:Animals; D000950:Antigens, Ly; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D055650:NK Cell Lectin-Like Receptor Subfamily A; D010750:Phosphoproteins / Q000276:immunology* / Q000378:metabolism; D011940:Receptor Aggregation; D011971:Receptors, Immunologic / Q000276:immunology* / Q000378:metabolism; D055630:Receptors, NK Cell Lectin-Like; D015398:Signal Transduction / Q000276:immunology*; D014443:Tyrosine
D000950:Antigens, Ly; D003432:Cross-Linking Reagents; C527092:Klra4 protein, mouse; D037181:Lectins, C-Type; D008562:Membrane Glycoproteins; D055650:NK Cell Lectin-Like Receptor Subfamily A; D010750:Phosphoproteins; D011971:Receptors, Immunologic; D055630:Receptors, NK Cell Lectin-Like; D014443:Tyrosine