Quantitative LC-MS/MS Analysis of Apical Membrane Proteome of mpkCCD Cells

This is a 2-plex SILAC-based quantitative proteomic study. It quantifies abundance changes of the apical membrane proteins of the kidney collecting duct model cells (mpkCCD) in response to vasopressin. Cells were labeled with normal or heavy lysine and arginine and then exposed to vehicle or the vasopressin analogy dDAVP (1 nM, 1 hour). Apical membrane proteins of vehicle and dDAVP-treated cells were enriched via surface biotinylation and affinity purification prior to LC-MS/MS analysis. Database search and protein quantification were performed using the SEQUEST algorithm included in the Proteome Discover Version 1.3 (Thermo Scientific). The database used for the spectral searching was prepared from the mouse RefSeq database plus common contaminants.