PSE132: Quantitative proteomic analysis of mpkCCD cells in response to long-term vasopressin exposure NCBI FTP Organism: Mus musculus Statistics Samples: View samples (5) Proteins: 4172 Peptides: 16124 Spectra: 165067 Dates Release: December 9, 2010 Deposit: January 27, 2010 Metadata update: December 10, 2010 Data update: January 27, 2010 Contact Organization: National Taiwan University College of Medicine Address: Rm 816, No. 1 Ren-Ai Road Section 1 Taipei 100 Overall Design Two mpkCCD cell populations were metabolically labeled with either light amino acids (C12 lysine or C12/N14 arginine) or heavy amino acids (C13 lysine and C13/N15 arginine). After complete labeling of the cells with isotopic amino acids, the mpkCCD cells were grown on membrane supports until confluence (transepithelial resistance > 5kohm x centimeter square) before they were exposed to 0.1nM dDAVP or vehicle for 5 days. Using two labeled cells (light and heavy amino acid labeled) combined with two conditions (dDAVP and vehicle exposures), proteins that change in abundances in response to dDAVP can be measured as a function of light to heavy peptide ratios. Summary Vaopressin is a key peptide hormone responsible for aquaporin 2 mRNA and protein expression in kidney collecting duct cells. Here we used SILAC (stable isotpic labelling by amino acids in cell culture) combined with LC-MS/MS (LTQ-FT) to quantify proteins that change in abundances in a collecting duct model cell line in response to a physiological concentration of vasopressin V2 receptor agonist, dDAVP, at 0.1nM for 5 days. Control cells were exposed to vehicle alone for the same period of time. The mpkCCD cells recapitulate vasopressin-mediated aquaporin 2 mRNA and protein expression as seen in native kidney collecting duct cells (Yu et al. PNAS 2009, 106(7): 2441-2446). Transcripts expressed in the mpkCCD cells can be found at the following link, http://dir.nhlbi.nih.gov/papers/lkem/mpkccdtr/Default.aspx. Contributors Ming-Jiun Yu Publications * Khositseth S, Pisitkun T, Slentz DH, Wang G et al. Quantitative protein and mRNA profiling shows selective post-transcriptional control of protein expression by vasopressin in kidney cells. Mol Cell Proteomics 2011 Jan;10(1):M110.004036. PMID: 20940332 Processing Methods Search Engines: OMSSA, SEQUEST, InsPecT Modifications Variable: Carbamidomethyl (4), Propionamide (24), Oxidation (35), Label:13C(6) (188), Label:13C(6)15N(4) (267) Platform(s) ESI-FTICR-CID