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The identification of cell surface accessible biomarkers enabling diagnosis, disease monitoring, and treatment of renalcell carcinoma (RCC) is as challenging as the biology and progression of RCC is unpredictable. A hallmark of most RCCis the loss-of-function of the von Hippel–Lindau (pVHL) protein by mutation of its gene (VHL). Using the cell surfacecapturing (CSC) technology, we screened and identified cell surface N-glycoproteins in pVHL-negative and positive786-O cells. One hundred six cell surface N-glycoproteins were identified. Stable isotope labeling with amino acidsin cell culture–based quantification of the CSC screen revealed 23 N-glycoproteins whose abundance seemed tochange in a pVHL-dependent manner. Targeted validation experiments using transcriptional profiling of primary RCCsamples revealed that nine glycoproteins, including CD10 and AXL, could be directly linked to pVHL-mediated transcrip-tional regulation. Subsequent human tumor tissue analysis of these cell surface candidate markers showed a correla-tion between epithelial AXL expression and aggressive tumor phenotype, indicating that pVHL-dependent regulation ofglycoproteins may influence the biologic behavior of RCC. Functional characterization of the metalloprotease CD10 incell invasion assays demonstrated a diminished penetrating behavior of pVHL-negative 786-O cells on treatment withthe CD10-specific inhibitor thiorphan. Our proteomic surfaceome screening approach in combination with transcrip-tional profiling and functional validation suggests pVHL-dependent cell surface glycoproteins as potential diagnosticmarkers for therapeutic targeting and RCC patient monitoring., Neoplasia, 14 (6), ISSN:1522-8002, ISSN:1476-5586