Alteration of nuclear protein profiling for NIH‐3T3 cells exposed to H2O2

Abstract

Oxidative stress is a threat to mammalian cells. To better understand the molecular response and mechanism underlying oxidative stress, we applied two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-fight mass spectrometry analysis to identify differential nuclear protein profiling of mouse fibroblast NIH-3T3 cells exposed to mild-H₂O₂. Thirteen differentially expressed proteins were identified by MS and two of them were further validated by Western blot. The results revealed that exposure to mild-H₂O₂ for 12 h cause up-regulated expression of DJ-1, glutathione S-transferase P 1, DNA ligase I, dynamin 2, nucleophosmin, and down-regulated expression of nucleoside diphosphate kinase A, enolase-α, barrier-to-autointegration factor 1, metastasis associated protein 1, glycosytransferase-like domain containing protein 1, synaptonemal complex protein 1, alpha-centractin, bromodomain, and PHD finger containing 1 (BRPF1). Most of the identified proteins are supported as nuclear proteins localized by previous research. The findings may provide some clues to elucidate cell responses to H₂O₂ and the potential mechanism underlying protection against oxidative stress in fibroblast cells.