Probing the folding intermediate of bacillus subtilis RNase P protein by nuclear magnetic resonance

Yu Chu Chang, William R. Franch, Terrence G. Oas

研究成果: 雜誌貢獻文章同行評審

8 引文 斯高帕斯(Scopus)

摘要

Protein folding intermediates are often imperative for overall folding processes and consequent biological functions. However, the low population and transient nature of the intermediate states often hinder their biochemical and biophysical characterization. Previous studies have demonstrated that Bacillus subtilis ribonuclease P protein (P protein) is conformationally heterogeneous and folds with multiphasic kinetics, indicating the presence of an equilibrium and kinetic intermediate in its folding mechanism. In this study, nuclear magnetic resonance (NMR) spectroscopy was used to study the ensemble corresponding to this intermediate (I). The results indicate that the N-terminal and C-terminal helical regions are mostly unfolded in I. 1H- 15N heteronuclear single-quantum coherence NMR spectra collected as a function of pH suggest that the protonation of His 22 may play a major role in the energetics of the equilibria among the unfolded, intermediate, and folded state ensembles of P protein. NMR paramagnetic relaxation enhancement experiments were also used to locate the small anion binding sites in both the intermediate and folded ensembles. The results for the folded protein are consistent with the previously modeled binding regions. These structural insights suggest a possible role for I in the RNase P holoenzyme assembly process.
原文英語
頁(從 - 到)9428-9437
頁數10
期刊Biochemistry
49
發行號44
DOIs
出版狀態已發佈 - 十一月 9 2010

ASJC Scopus subject areas

  • 生物化學

指紋

深入研究「Probing the folding intermediate of bacillus subtilis RNase P protein by nuclear magnetic resonance」主題。共同形成了獨特的指紋。

引用此