Project PXD000183



Human kidney primary proximal tubule cell LC-MSMS


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.

Sample Processing Protocol

See details in reference(s) : 23846697

Data Processing Protocol

See details in reference(s) : 23846697


Ana Konvalinka, Department of the Institute of Medical Sciences, Division of Nephrology

Submission Date


Publication Date



Not available


LTQ Orbitrap


Not available

Experiment Type

Bottom-up proteomics


    Konvalinka A, Zhou J, Dimitromanolakis A, Drabovich AP, Fang F, Gurley S, Coffman T, John R, Zhang SL, Diamandis EP, Scholey JW; Determination of an Angiotensin II-regulated Proteome in Primary Human Kidney Cells by Stable Isotope Labeling of Amino Acids in Cell Culture (SILAC)., J Biol Chem, 2013 Aug 23, 288, 34, 24834-47, PubMed: 23846697