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

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

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)9428-9437
Number of pages10
JournalBiochemistry
Volume49
Issue number44
DOIs
Publication statusPublished - Nov 9 2010
Externally publishedYes

Fingerprint

Ribonuclease P
Bacilli
Bacillus subtilis
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Proteins
Biological Phenomena
Protein folding
Holoenzymes
Kinetics
Protonation
Protein Folding
Nuclear magnetic resonance spectroscopy
Anions
Binding Sites
Population
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

Probing the folding intermediate of bacillus subtilis RNase P protein by nuclear magnetic resonance. / Chang, Yu Chu; Franch, William R.; Oas, Terrence G.

In: Biochemistry, Vol. 49, No. 44, 09.11.2010, p. 9428-9437.

Research output: Contribution to journalArticle

Chang, Yu Chu ; Franch, William R. ; Oas, Terrence G. / Probing the folding intermediate of bacillus subtilis RNase P protein by nuclear magnetic resonance. In: Biochemistry. 2010 ; Vol. 49, No. 44. pp. 9428-9437.
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