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The Journal of Immunology, 1998, 160: 4148-4152.
Copyright © 1998 by The American Association of Immunologists


CUTTING EDGE

Cutting Edge: Characterization of an Associated 16-kDa Tyrosine Phosphoprotein Required for Ly-49D Signal Transduction3

Llewellyn H. Mason4,*, Jami Willette-Brown*, Stephen K. Anderson{dagger}, Pierre Gosselin*, Elizabeth W. Shores{ddagger}, Paul E. Love§, John R. Ortaldo* and Daniel W. McVicar*

* Laboratory of Experimental Immunology, Division of Basic Sciences, and {dagger} Intramural Research Support Program, SAIC Frederick, National Cancer Institute-Frederick Cancer Reseach and Development Center, Frederick, MD 21702; {ddagger} Division of Hematologic Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Washington, DC; and § Laboratory of Mammalian Genes and Development, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892

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{epsilon}RI{gamma} double knockout mice confirm that pp16 is not TCR-{zeta}, TCR-{eta}, or Fc{epsilon}RI{gamma}. Association of pp16 with Ly-49D involves a transmembrane arginine since mutation to leucine (Ly-49DR54L) abolishes association with pp16 in transfected P815 cells. In addition, Ly-49DR54L transfectants fail to mediate Ca2+ mobilization following Ab cross-linking. Therefore, signaling through Ly-49D 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.




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