Quantitative Phosphoproteomics of CXCL12 (SDF-1) Signaling 英文参考文献.docVIP

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Quantitative Phosphoproteomics of CXCL12 (SDF-1) Signaling 英文参考文献.doc

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Quantitative Phosphoproteomics of CXCL12 (SDF-1) Signaling 英文参考文献

QuantitativePhosphoproteomicsofCXCL12(SDF-1) Signaling JasonA.Wojcechowskyj1,JessicaY.Lee2,StevenH.Seeholzer2,RobertW.Doms1* 1Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America, 2Protein and Proteomics Core, Children’sHospitalofPhiladelphiaResearchInstitute,Philadelphia,Pennsylvania,UnitedStatesofAmerica Abstract CXCL12(SDF-1)isachemokinethatbindstoandsignalsthroughtheseventransmembranereceptorCXCR4.TheCXCL12/ CXCR4 signaling axis has been implicated in both cancer metastases and human immunodeficiency virus type 1 (HIV-1) infection and a more complete understanding of CXCL12/CXCR4 signaling pathways may support efforts to develop therapeutics for these diseases. Mass spectrometry-based phosphoproteomics has emerged as an important tool in studyingsignalingnetworksinanunbiasedfashion.Weemployedstableisotopelabelingwithaminoacidsincellculture (SILAC)quantitativephosphoproteomicstoexaminetheCXCL12/CXCR4signalingaxisinthehumanlymphoblasticCEMcell line.Wequantified4,074uniqueSILACpairsfrom1,673proteinsand89phosphopeptidesweredeemedCXCL12-responsive in biological replicates. Several well established CXCL12-responsive phosphosites such as AKT (pS473) and ERK2 (pY204) wereconfirmedinourstudy.WealsovalidatedtwonovelCXCL12-responsivephosphosites,stathmin(pS16)andAKT1S1 (pT246) by Westernblot. Pathwayanalysis and comparisons with otherphosphoproteomic datasetsrevealedthat genes fromCXCL12-responsivephosphositesareenrichedforcellularpathwayssuchasTcellactivation,epidermalgrowthfactor and mammalian target of rapamycin (mTOR) signaling, pathways which have previously been linked to CXCL12/CXCR4 signaling.SeveralofthenovelCXCL12-responsivephosphoproteinsfromourstudyhavealsobeenimplicatedwithcellular migrationandHIV-1infection,thusprovidinganattractivelistofpotentialtargetsforthedevelopmentofcancermetastasis and HIV-1 therapeutics and for furthering our understanding of chemokine signaling regulation