Screening of differential proteins binding to Nox1 promoter in A549 cell model of inflammation and oxidative stress.
- Author:
Xian QIU
1
;
Shuiwang HU
;
Jun XU
;
Li LI
;
Wenjie HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; DEAD-box RNA Helicases; metabolism; Electrophoresis, Gel, Two-Dimensional; Humans; Inflammation; Keratin-1; metabolism; Keratin-10; metabolism; Mass Spectrometry; NADPH Oxidase 1; NADPH Oxidases; genetics; metabolism; Nucleocytoplasmic Transport Proteins; metabolism; Oxidative Stress; Promoter Regions, Genetic; Tumor Necrosis Factor-alpha; adverse effects
- From: Journal of Southern Medical University 2013;33(5):703-707
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen the regulatory proteins involved in Nox1 promoter activation in a cell model of inflammation and oxidative stress.
METHODSA cell model of inflammation and oxidative stress was established by stimulating A549 cells with tumor necrosis factor-α (TNF-α). The differential proteins binding to Nox1 promoter were screened by DNA pull-down and the binding proteins were separated by 2D electrophoresis and selected according to the their differential expression levels (with over 1.5-fold changes relative to the control level). The screened proteins were finally identified by MALDI-TOF/TOF-MS.
RESULTSSeven differentially expressed protein spots (all upregulated in the cell model) were obtained, among which GLE1, DDX19A, KRT1 and KRT10 were identified by mass spectrometry.
CONCLUSIONGLE1, DDX19A, KRT1 and KRT10 participate in the activation of Nox1 promoter in TNF-α-induced A549 cells, and this result provides new insights into the biological roles of the regulatory proteins of Nox1 promoter in inflammation and oxidative stress.
