Journal article
Combinatorial proteomic analysis of intercellular signaling applied to the CD28 T-cell costimulatory receptor
Proceedings of the National Academy of Sciences - PNAS, Vol.112(13), pp.E1594-E1603
31/Mar/2015
PMCID: PMC4386406
PMID: 25829543
Abstract
Systematic characterization of intercellular signaling approximating the physiological conditions of stimulation that involve direct cell-cell contact is challenging. We describe a proteomic strategy to analyze physiological signaling mediated by the T-cell costimulatory receptor CD28. We identified signaling pathways activated by CD28 during direct cell-cell contact by global analysis of protein phosphorylation. To define immediate CD28 targets, we used phosphorylated forms of the CD28 cytoplasmic region to obtain the CD28 interactome. The interaction profiles of selected CD28-interacting proteins were further characterized in vivo for amplifying the CD28 interactome. The combination of the global phosphorylation and interactome analyses revealed broad regulation of CD28 and its interactome by phosphorylation. Among the cellular phosphoproteins influenced by CD28 signaling, CapZ-interacting protein (CapZIP), a regulator of the actin cytoskeleton, was implicated by functional studies. The combinatorial approach applied herein is widely applicable for characterizing signaling networks associated with membrane receptors with short cytoplasmic tails.
Details
- Title
- Combinatorial proteomic analysis of intercellular signaling applied to the CD28 T-cell costimulatory receptor
- Creators
- Ruijun Tian (Corresponding Author) - Lunenfeld-Tanenbaum Research InstituteHaopeng Wang - University of California, San FranciscoGerald D Gish - Lunenfeld-Tanenbaum Research InstituteEvangelia Petsalaki - Lunenfeld-Tanenbaum Research InstituteAdrian Pasculescu - Lunenfeld-Tanenbaum Research InstituteYu Shi - Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, CA 92037Marianne Mollenauer - University of California, San FranciscoRichard D Bagshaw - Lunenfeld-Tanenbaum Research InstituteNir Yosef - University of California, BerkeleyTony Hunter - Salk Institute for Biological StudiesAnne-Claude Gingras - University of TorontoArthur Weiss (Corresponding Author) - University of California, San FranciscoTony Pawson
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.112(13), pp.E1594-E1603; 31/Mar/2015
- Number of pages
- 10
- Language
- English
- DOI
- https://doi.org/10.1073/pnas.1503286112
- PMID
- 25829543
- PMCID
- PMC4386406
- Grant note
- We acknowledge K. Colwill, L. Taylor, B. Larsen, C. Zhang, and M. Tucholska for technical help and J. Jin, Y. Zheng, C. Chen, G. M. Findlay, and M. Kofler for helpful discussions. We thank A. Salomon and N. Krogan for constructive suggestions after their reading of this manuscript. We thank J. Bluestone, who provided the CTLA4-Ig. This work was supported by grants from Ontario Research Fund GL2 (to T.P. and A.-C.G.), the Canadian Institutes of Health Research (CIHR) (Grant MOP-84314) (to A.-C.G.), the Howard Hughes Medical Institute (to A.W.), the NIH/National Cancer Institute (Grant CA82683) (to T.H.), and a Shenzhen grant (ZDSYS20140509142721429) (to R.T.). R.T. is the recipient of a fellowship from the CIHR. H.W. is the recipient of a postdoctoral fellowship from the US Arthritis Foundation. Author contributions: R.T., H.W., A.W., and T.P. designed research; R.T., H.W., G.D.G., E.P.,A.P., Y.S., M.M., and R.D.B. performed research; N.Y., T.H., and A.-C.G. contributed new reagents/analytic tools; R.T., H.W., G.D.G., E.P., A.P., Y.S., M.M., R.D.B., A.W., and T. P. analyzed data; and R.T., H.W., A.W., and T.P. wrote the paper.
- Record Identifier
- 993427183203596
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