Proteomics

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Mass spectrometry analysis of in vitro kinase assay for DNA-PKcs and TAF7


ABSTRACT: To examine whether TAF7 could be phosphorylated by DNA-PKcs directly. An in vitro kinase assays were performed. Firstly, human DNA-PKcs was purified from TGF1 treated HK2 cells by immunoprecipitation with a DNA-PKcs antibody (abcam) and IgG was used as a negative control. The purified proteins and human GST-TAF7 (purchased from proteintech) were incubated in kinase assay buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 10 mM MgCl2, and 1 mM MnCl2) supplemented with 200 M ATP in 50 l reactions for 30 min at 30°C. Then, phosphorylation of TAF7 was analyzed by mass spectrometry (MS)

ORGANISM(S): Homo Sapiens

SUBMITTER: Zhanjun Jia  

PROVIDER: PXD036930 | iProX | Thu Sep 22 00:00:00 BST 2022

REPOSITORIES: iProX

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Publications

DNA-dependent protein kinase catalytic subunit (DNA-PKcs) drives chronic kidney disease progression in male mice.

Yang Yunwen Y   Liu Suwen S   Wang Peipei P   Ouyang Jing J   Zhou Ning N   Zhang Yue Y   Huang Songming S   Jia Zhanjun Z   Zhang Aihua A  

Nature communications 20230311 1


Kidney injury initiates epithelial dedifferentiation and myofibroblast activation during the progression of chronic kidney disease. Herein, we find that the expression of DNA-PKcs is significantly increased in the kidney tissues of both chronic kidney disease patients and male mice induced by unilateral ureteral obstruction and unilateral ischemia-reperfusion injury. In vivo, knockout of DNA-PKcs or treatment with its specific inhibitor NU7441 hampers the development of chronic kidney disease in  ...[more]

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