title: GRID: a novel Grb-2-related adapter protein that interacts with the activated T cell costimulatory receptor CD28.
abstract: Adapter proteins such as Grb2 play a central role in the formation of signaling complexes through their association with multiple protein binding partners. These interactions are mediated by specialized domains such as the well-characterized Src homology SH2 and SH3 motifs. Using yeast three-hybrid technology, we have identified a novel adapter protein, expressed predominantly in T lymphocytes, that associates with the activated form of the costimulatory receptor, CD28. The protein is a member of the Grb2 family of adapter proteins and contains an SH3-SH2-SH3 domain structure. A unique glutamine/proline-rich domain (insert domain) of unknown function is situated between the SH2 and N-terminal SH3 domains. We term this protein GRID for Grb2-related protein with insert domain. GRID coimmunoprecipitates with CD28 from Jurkat cell lysates following activation of CD28. Using mutants of CD28 and GRID, we demonstrate that interaction between the proteins is dependent on phosphorylation of CD28 at tyrosine 173 and integrity of the GRID SH2 domain, although there are also subsidiary stabilizing contacts between the PXXP motifs of CD28 and the GRID C-terminal SH3 domain. In addition to CD28, GRID interacts with a number of other T cell signaling proteins, including SLP-76 (SH2 domain-containing leukocyte protein of 76 kDa), p62dok, and RACK-1 (receptor for activated protein kinase C-1). These findings suggest that GRID functions as an adapter protein in the CD28-mediated costimulatory pathway in T cells.
authors.affiliation: Immunopathology and Immunology Units, GlaxoWellcome Medicines Research Centre, Stevenage, United Kingdom. jhe45655@glaxowellcome.co.uk
authors.forename: J H
authors.initials: JH
authors.lastname: Ellis
authors.forename: C
authors.initials: C
authors.lastname: Ashman
authors.forename: M N
authors.initials: MN
authors.lastname: Burden
authors.forename: K E
authors.initials: KE
authors.lastname: Kilpatrick
authors.forename: M A
authors.initials: MA
authors.lastname: Morse
authors.forename: P A
authors.initials: PA
authors.lastname: Hamblin
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D000818:Animals; D018106:CD28 Antigens / Q000378:metabolism*; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D006801:Humans; D019169:Jurkat Cells; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis; D015398:Signal Transduction / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D018909:src Homology Domains / Q000235:genetics / Q000276:immunology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018106:CD28 Antigens; D002352:Carrier Proteins; C409587:GRAP2 protein, human; C498726:Mona protein, mouse; D012333:RNA, Messenger