Proteomics

Dataset Information

0

Identification of PKC-alpha dependent phosphoproteins in mouse retina


ABSTRACT: Adjusting to a wide range of light intensities is an essential feature of retinal rod bipolar cell (RBC) function, and persuasive evidence suggests this modulation involves phosphorylation by protein kinase C-alpha (PKC-alpha). PKC-alpha is a serine/threonine kinase that is strongly expressed in RBCs, but the targets of PKC-alpha phosphorylation in the retina have not been identified. PKC-alpha activity and phosphorylation in RBCs was examined by immunofluorescence confocal microscopy using a conformation-specific PKC-alpha antibody and antibodies to phosphorylated PKC motifs. PKC-alpha activity was dependent on light and expression of TRPM1, and RBC dendrites were the primary sites of light-dependent phosphorylation. PKC-alpha-dependent retinal phosphoproteins were identified using a multiplexed tandem mass tag-based approach to compare total protein and phosphopeptide abundance between phorbol ester-treated wild type and PKC-alpha knockout (PKC-alpha-KO) mouse retinas. Of the 23 proteins showing significant differential abundance between the two groups, most have roles in cytoskeleton/transport, transcriptional regulation, or metabolism/homeostasis. Phosphopeptide mass spectrometry identified over 1100 phosphopeptides in mouse retina, with 12 displaying significantly greater phosphorylation in WT compared to PKC-alpha-KO samples. The differentially phosphorylated proteins fall into the following functional groups: cytoskeleton/transport (4 proteins), ECM/adhesion (2 proteins), signaling (2 proteins), transcriptional regulation (3 proteins), and homeostasis/metabolism (1 protein). Two strongly differentially expressed phosphoproteins, BORG4 and TPBG, were localized to the synaptic layers of the retina, and may play a role in PKC-alpha-dependent modulation of RBC physiology.

INSTRUMENT(S): Orbitrap Fusion ETD

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Retinal Rod Cell, Retina

SUBMITTER: Phillip Wilmarth  

LAB HEAD: Dr. Catherine Morgans

PROVIDER: PXD012906 | Pride | 2019-07-15

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
GOTerms_report_WP.txt Txt
GOTerms_report_phospho.txt Txt
Log2R_peptides.xlsx Xlsx
Log2R_plotter.xls Xls
Log2R_proteins.xlsx Xlsx
Items per page:
1 - 5 of 80
altmetric image

Publications

Identification of PKCα-dependent phosphoproteins in mouse retina.

Wakeham Colin M CM   Wilmarth Phillip A PA   Cunliffe Jennifer M JM   Klimek John E JE   Ren Gaoying G   David Larry L LL   Morgans Catherine W CW  

Journal of proteomics 20190628


Adjusting to a wide range of light intensities is an essential feature of retinal rod bipolar cell (RBC) function. While persuasive evidence suggests this modulation involves phosphorylation by protein kinase C-alpha (PKCα), the targets of PKCα phosphorylation in the retina have not been identified. PKCα activity and phosphorylation in RBCs was examined by immunofluorescence confocal microscopy using a conformation-specific PKCα antibody and antibodies to phosphorylated PKC motifs. PKCα activity  ...[more]

Similar Datasets

2023-10-09 | PXD039490 | Pride
2022-02-17 | PXD026764 | Pride
2014-01-13 | E-GEOD-52006 | biostudies-arrayexpress
2012-11-02 | E-GEOD-41821 | biostudies-arrayexpress
2023-08-16 | PXD044270 | Pride
2023-08-16 | PXD044269 | Pride
2009-05-31 | E-GEOD-12601 | biostudies-arrayexpress
2014-09-10 | E-GEOD-59909 | biostudies-arrayexpress
2014-07-30 | E-GEOD-56563 | biostudies-arrayexpress
2021-09-10 | PXD023704 | Pride