Transcript Powerpoint
Cell surface receptors Ly-9 and CD84 recruit the X-linked lymphoproliferative disease gene product SAP by Joan Sayós, Margarita Martı́n, Alice Chen, Marı́a Simarro, Duncan Howie, Massimo Morra, Pablo Engel, and Cox Terhorst Blood Volume 97(12):3867-3874 June 15, 2001 ©2001 by American Society of Hematology The cell surface molecule 2B4 recruits the XLP gene product SAP after phosphorylation of the tyrosine residues in its cytoplasmic tail. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology SAP interacts with the phosphorylated cytoplasmic tail of Ly-9.A human cDNA library made from poly A+ RNA of the human T-cell line KT3 in pGAD424 was screened with the altered yeast 2hybrid system. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology Interactions of SAP with the phosphorylated cytoplasmic tail of CD84.(A) CD84 and SAP in yeast. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology Schematic representation of the cytoplasmic domains of 4 SLAM family members.SAP has been shown to bind preferentially to a consensus binding motif: T (I/V)Y x x (V/I), where x represents any amino acid. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology SAP blocks binding of SHP-2 to phosphorylated Ly-9 and CD84.To detect interactions between Ly-9 or CD84 and SHP-2, COS-7 cells were cotransfected with cDNAs encoding one of the receptors, humanc-fyn, in the presence or absence of human SAP. On immunoprecipi... Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology Phosphorylated CD84 and Ly-9 associate with SAP at the plasma membrane.To verify the results obtained with the yeast 2-hybrid system and immunoprecipitation in COS-7 cells, a fluorescence microscopy approach was used. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology CD84 and Ly-9 colocalize with SAP in Jurkat after activation of the receptors with monoclonal antibody.A fluorescence microscopy approach was used in the human T-cell line Jurkat to confirm the SAP/Ly-9 and SAP/CD84 interaction in T cells. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology Summary of the SAP-binding proteins.Binding of SAP to the cytoplasmic tail of the cell surface receptors is indicated by a plus sign, and absence of binding to SAP is indicated by a minus sign. Joan Sayós et al. Blood 2001;97:3867-3874 ©2001 by American Society of Hematology