Proteomics

Dataset Information

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Human kidney primary proximal tubule cell LC-MSMS


ABSTRACT: Angiotensin II (AngII), the effector of the renin angiotensin system causes kidney disease progression by signalling through AT-1 receptor, but there are no measures of AngII activity in the kidney. Accordingly, we sought to define an AngII-regulated proteome in primary human proximal tubular epithelial cells (PTEC) in order to identify potential markers of AngII activity in the kidney. PTECs were labeled with SILAC heavy arginine(+6) and lysine(+8) OR light arginine and lysine for 6 doubling times. Heavy-labeled PTECs were stimulated with AngII and light-labeled PTECs were stimulated with the control medium. AngII-treated and control-treated PTEC lysates were mixed in 1:1 total protein ratio and their proteomes were compared. We generated 5 biological replicates (1 supernatant, 1 replicate with reverse labeling). LTQ-Orbitrap mass spectrometer was used for LC-MSMS analysis. Of 4618 quantified proteins, 83 were differentially regulated by AngII in 4 biological replicates of PTEC lysates. We subsequently confirmed and verified 18 differentially regulated proteins by using SRM, RT-PCR and ELISA. We also went on to validate the main functional, enriched networks by using systems biology approach and an in vivo mouse model.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Ana Konvalinka  

PROVIDER: PXD000183 | Pride | 2013-07-18

REPOSITORIES: Pride

Dataset's files

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Publications

Determination of an angiotensin II-regulated proteome in primary human kidney cells by stable isotope labeling of amino acids in cell culture (SILAC).

Konvalinka Ana A   Zhou Joyce J   Dimitromanolakis Apostolos A   Drabovich Andrei P AP   Fang Fei F   Gurley Susan S   Coffman Thomas T   John Rohan R   Zhang Shao-Ling SL   Diamandis Eleftherios P EP   Scholey James W JW  

The Journal of biological chemistry 20130711 34


Angiotensin II (AngII), the major effector of the renin-angiotensin system, mediates kidney disease progression by signaling through the AT-1 receptor (AT-1R), but there are no specific measures of renal AngII activity. Accordingly, we sought to define an AngII-regulated proteome in primary human proximal tubular cells (PTEC) to identify potential AngII activity markers in the kidney. We utilized stable isotope labeling with amino acids (SILAC) in PTECs to compare proteomes of AngII-treated and  ...[more]

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