quantitative phosphoproteomics of cxcl12 (sdf-1) signaling定量phosphoproteomics cxcl12(sdf-1)信号.pdfVIP

quantitative phosphoproteomics of cxcl12 (sdf-1) signaling定量phosphoproteomics cxcl12(sdf-1)信号.pdf

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quantitative phosphoproteomics of cxcl12 (sdf-1) signaling定量phosphoproteomics cxcl12(sdf-1)信号

Quantitative Phosphoproteomics of CXCL12 (SDF-1) Signaling 1 2 2 1 Jason A. Wojcechowskyj , Jessica Y. Lee , Steven H. Seeholzer , Robert W. Doms * 1 Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, United States of America, 2 Protein and Proteomics Core, Children’s Hospital of Philadelphia Research Institute, Philadelphia, Pennsylvania, United States of America Abstract CXCL12 (SDF-1) is a chemokine that binds to and signals through the seven transmembrane receptor CXCR4. The CXCL12/ 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 studying signaling networks in an unbiased fashion. We employed stable isotope labeling with amino acids in cell culture (SILAC) quantitative phosphoproteomics to examine the CXCL12/CXCR4 signaling axis in the human lymphoblastic CEM cell line. We quantified 4,074 unique SILAC pairs from 1,673 proteins and 89 phosphopeptides were deemed CXCL12-responsive in biological replicates. Several well established CXCL12-responsive phosphosites such as AKT (pS473) and ERK2 (pY204) were confirmed in our study. We also validated two novel CXCL12-responsive phosphosites, stathmin (pS16) and AKT1S1 (pT246) by Western blot. Pathway analysis and comparisons with other phosphoproteomic datasets revealed that genes from CXCL12-responsive phosphosites are enriched for cellular pathways such as T cell activation, epidermal growth factor an

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