Quantitative phosphoproteomic analysis reveals vasopressin V2-receptor-dependent signaling pathways in renal collecting duct cells

被引:139
作者
Rinschen, Markus M. [1 ,2 ]
Yu, Ming-Jiun [1 ]
Wang, Guanghui [3 ]
Boja, Emily S. [3 ]
Hoffert, Jason D. [1 ]
Pisitkun, Trairak [1 ]
Knepper, Mark A. [1 ]
机构
[1] NHLBI, Epithelial Syst Biol Lab, NIH, Bethesda, MD 20892 USA
[2] Univ Munster, Dept Internal Med D, Munster, Germany
[3] NHLBI, Prote Core Facil, NIH, Bethesda, MD 20892 USA
关键词
aquaporin-2; MAP kinase; mass spectrometry; protein kinase A; SILAC; PHOSPHORYLATION ANALYSIS; MASS-SPECTROMETRY; KINASE-II; AQUAPORIN-2; KIDNEY; EXPRESSION; IDENTIFICATION; PEPTIDES; TARGET; TOOL;
D O I
10.1073/pnas.0910646107
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Vasopressin's action in renal cells to regulate water transport depends on protein phosphorylation. Here we used mass spectrometry-based quantitative phosphoproteomics to identify signaling pathways involved in the short-term V2-receptor-mediated response in cultured collecting duct cells (mpkCCD) from mouse. Using Stable Isotope Labeling by Amino acids in Cell culture (SILAC) with two treatment groups (0.1 nM dDAVP or vehicle for 30 min), we carried out quantification of 2884 phosphopeptides. The majority (82%) of quantified phosphopeptides did not change in abundance in response to dDAVP. Analysis of the 273 phosphopeptides increased by dDAVP showed a predominance of so-called "basophilic" motifs consistent with activation of kinases of the AGC family. Increases in phosphorylation of several known protein kinase A targets were found. In addition, increased phosphorylation of targets of the calmodulin-dependent kinase family was seen, including autophosphorylation of calmodulin-dependent kinase 2 at T286. Analysis of the 254 phosphopeptides decreased in abundance by dDAVP showed a predominance of so-called "proline-directed" motifs, consistent with down-regulation of mitogen-activated or cyclin-dependent kinases. dDAVP decreased phosphorylation of both JNK1/2 (T183/Y185) and ERK1/2 (T183/Y185; T203/Y205), consistent with a decrease in activation of these proline-directed kinases in response to dDAVP. Both ERK and JNK were able to phosphorylate residue S261 of aquaporin-2 in vitro, a site showing a decrease in phosphorylation in response to dDAVP in vivo. The data support roles for multiple vasopressin V2-receptor-dependent signaling pathways in the vasopressin signaling network of collecting duct cells, involving several kinases not generally accepted to regulate collecting duct function.
引用
收藏
页码:3882 / 3887
页数:6
相关论文
共 35 条
  • [1] Identification of c-Src tyrosine kinase substrates in platelet-derived growth factor receptor signaling
    Amanchy, Ramars
    Zhong, Jun
    Hong, Rosa
    Kim, James H.
    Gucek, Marjan
    Cole, Robert N.
    Molina, Henrik
    Pandey, Akhilesh
    [J]. MOLECULAR ONCOLOGY, 2009, 3 (5-6): : 439 - 450
  • [2] A probability-based approach for high-throughput protein phosphorylation analysis and site localization
    Beausoleil, Sean A.
    Villen, Judit
    Gerber, Scott A.
    Rush, John
    Gygi, Steven P.
    [J]. NATURE BIOTECHNOLOGY, 2006, 24 (10) : 1285 - 1292
  • [3] BRICKEY DA, 1994, J BIOL CHEM, V269, P29047
  • [4] Phosphorylation events and the modulation of aquaporin 2 cell surface expression
    Brown, Dennis
    Hasler, Udo
    Nunes, Paula
    Bouley, Richard
    Lu, Hua A. J.
    [J]. CURRENT OPINION IN NEPHROLOGY AND HYPERTENSION, 2008, 17 (05) : 491 - 498
  • [5] Chou C.-L., 1999, Journal of the American Society of Nephrology, V10, p13A
  • [6] Non-muscle myosin II and myosin light chain kinase are downstream targets for vasopressin signaling in the renal collecting duct
    Chou, CL
    Christensen, BM
    Frische, S
    Vorum, H
    Desai, RA
    Hoffert, JD
    de Lanerolle, P
    Nielsen, S
    Knepper, MA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (47) : 49026 - 49035
  • [7] Signal transduction by the JNK group of MAP kinases
    Davis, RJ
    [J]. CELL, 2000, 103 (02) : 239 - 252
  • [8] Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry
    Elias, Joshua E.
    Gygi, Steven P.
    [J]. NATURE METHODS, 2007, 4 (03) : 207 - 214
  • [9] AN APPROACH TO CORRELATE TANDEM MASS-SPECTRAL DATA OF PEPTIDES WITH AMINO-ACID-SEQUENCES IN A PROTEIN DATABASE
    ENG, JK
    MCCORMACK, AL
    YATES, JR
    [J]. JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 1994, 5 (11) : 976 - 989
  • [10] Open mass spectrometry search algorithm
    Geer, LY
    Markey, SP
    Kowalak, JA
    Wagner, L
    Xu, M
    Maynard, DM
    Yang, XY
    Shi, WY
    Bryant, SH
    [J]. JOURNAL OF PROTEOME RESEARCH, 2004, 3 (05) : 958 - 964