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Global proteomic profiling of vasopressin‐dependent nuclear localization
Author(s) -
Schenk Laura Katharina,
Pisitkun Trairak,
Miller Lance,
Sandoval Pablo,
Hoffert Jason,
Knepper Mark
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb644
Subject(s) - stable isotope labeling by amino acids in cell culture , chemistry , vasopressin , microbiology and biotechnology , gene expression , isobaric labeling , nuclear transport , compartment (ship) , cytoplasm , quantitative proteomics , nuclear protein , transcription factor , proteomics , gene , biology , biochemistry , cell nucleus , endocrinology , oceanography , geology
Vasopressin regulates the expression of many genes in the renal collecting duct. To identify mediators, we have carried out LC‐MS/MS profiling of cytoplasm, soluble nuclear compartment, and nuclear pellet of collecting duct mpkCCD cells. Stable isotope labeling (SILAC) was used for relative quantification of tryptic peptides in vasopressin‐treated (1h dDAVP, 0.1nM) compared to control cells. There were no measurable differences in total protein content. We identified 5776 peptides corresponding to 1074 proteins in the soluble nuclear compartment. 49 proteins were either decreased or increased in abundance (SILAC isotope ratio ≤0.7 or ≥1.3). In the nuclear pellet, we identified 57749 peptides (1383 proteins). 224 of these showed altered abundance in presence of dDAVP. Altered proteins are candidates to play a regulatory role on gene expression upon dDAVP treatment, e.g. regarding aquaporin‐2, whether changes are due to nuclear translocation or post‐translational modification. One example is the CREB‐regulated transcription coactivator 1 (Crtc1) protein, which is increased in the soluble nuclear compartment in response to vasopressin.