Proteomics

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

ABSTRACT: 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.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Ming-Jiun Yu  

LAB HEAD: Ming-Jiun Yu

PROVIDER: PXD000399 | Pride | 2013-10-02

REPOSITORIES: Pride

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Dataset's files

Source:
Action DRS
1st_SILAC_Data.xlsx Xlsx
1st_SILAC_Search.msf Msf
1st_SILAC_pep.xml Xml
2nd_SILAC_Data.xlsx Xlsx
2nd_SILAC_Search.msf Msf
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Publications

Quantitative apical membrane proteomics reveals vasopressin-induced actin dynamics in collecting duct cells.

Loo Chin-San CS   Chen Cheng-Wei CW   Wang Po-Jen PJ   Chen Pei-Yu PY   Lin Shu-Yu SY   Khoo Kay-Hooi KH   Fenton Robert A RA   Knepper Mark A MA   Yu Ming-Jiun MJ  

Proceedings of the National Academy of Sciences of the United States of America 20131001 42

In kidney collecting duct cells, filamentous actin (F-actin) depolymerization is a critical step in vasopressin-induced trafficking of aquaporin-2 to the apical plasma membrane. However, the molecular components of this response are largely unknown. Using stable isotope-based quantitative protein mass spectrometry and surface biotinylation, we identified 100 proteins that showed significant abundance changes in the apical plasma membrane of mouse cortical collecting duct cells in response to vas  ...[more]

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