Oxidation-Induced Conformational Change of a Prokaryotic Molecular Chaperone, Hsp33, Monitored by Selective Isotope Labeling using 1H NMR spectroscopy coupled with multivariate analysis
2
조회
0
다운로드
6
인용
0
참고문헌
키워드
초록
Hsp33, a prokaryotic molecular chaperone, exerts holdase activity in response to oxidative stress. In this study, the stepwise conformational change of Hsp33 upon oxidation was monitored by NMR. In order to overcome its high molecular weight (33 kDa as a monomer and 66 kDa as a dimer), spectra were simplified using a selectively [^(15)N]His-labeled protein. All of the eight histidines were observed in the TROSY spectrum of the reduced Hsp33. Among them, three peaks showed dramatic resonance shifts dependent on the stepwise oxidation, indicating a remarkable conformational change. The results suggest that unfolding of the linker domain is associated with dimerization, but not entire region of the linker domain is unfolded.
Abstract
Hsp33, a prokaryotic molecular chaperone, exerts holdase activity in response to oxidative stress. In this study, the stepwise conformational change of Hsp33 upon oxidation was monitored by NMR. In order to overcome its high molecular weight (33 kDa as a monomer and 66 kDa as a dimer), spectra were simplified using a selectively [^(15)N]His-labeled protein. All of the eight histidines were observed in the TROSY spectrum of the reduced Hsp33. Among them, three peaks showed dramatic resonance shifts dependent on the stepwise oxidation, indicating a remarkable conformational change. The results suggest that unfolding of the linker domain is associated with dimerization, but not entire region of the linker domain is unfolded.
참고문헌 (14)
- U. Jakob, , Cell, 96, pp.341~ (1999)
- P.C.F. Graf, , Cell. Mol. Life Sci, 59, pp.1624~ (2002)
- M.W. Akhtar, , J. Biol. Chem, 279, pp.55760~ (2004)
- M. Ilbert, , Nat. Struct. Mol. Biol, 14, pp.556~ (2007)
- J. Graumann, , Structure, 9, pp.377~ (2001)
- P.C.F. Graf, , J. Bio. Chem, 279, pp.20529~ (2004)
- I. Janda, , Structure, 12, pp.1901~ (2004)
- J. Vijayalakshmi, , Structure, 9, pp.367~ (2001)
- S.-J. Kim, , Nat. Struct. Biol, 8, pp.459~ (2001)
- L. Jaroszewski, , Proteins, 61, pp.669~ (2005)
- S-W. Chi, , FEBS Lett, 585, pp.664~ (2011)
- labeling and NMR measurements of the inactive, reduced monomer form of Escherichia coli Hsp33, "Journal of the Korean Magnetic Resonance Society", 2010, 117~126, pp.한국자기공명학회
- M. Ilbert, , Antioxid. Redox Signal, 8, pp.835~ (2006)
- Dae-Won Sim, "Backbone NMR assignments of a putative secretory protein from Helicobacter pylori, using ahigh - field (900MHz) NMR", Journal of the Korean Magnetic Resonance Society, 13(2), pp.108~116 (2009), 한국자기공명학회