Reversible unfolding of a thermophilic membrane protein in phospholipid/detergent mixed micelles

Journal of Molecular Biology
Ernesto A RomanF L González Flecha

Abstract

Folding mechanisms and stability of membrane proteins are poorly understood because of the known difficulties in finding experimental conditions under which reversible denaturation could be possible. In this work, we describe the equilibrium unfolding of Archaeoglobus fulgidus CopA, an 804-residue alpha-helical membrane protein that is involved in transporting Cu(+) throughout biological membranes. The incubation of CopA reconstituted in phospholipid/detergent mixed micelles with high concentrations of guanidinium hydrochloride induced a reversible decrease in fluorescence quantum yield, far-UV ellipticity, and loss of ATPase and phosphatase activities. Refolding of CopA from this unfolded state led to recovery of full biological activity and all the structural features of the native enzyme. CopA unfolding showed typical characteristics of a two-state process, with DeltaG(w) degrees =12.9 kJ mol(-)(1), m=4.1 kJ mol(-1) M(-1), C(m)=3 M, and DeltaCp(w) degrees =0.93 kJ mol(-1) K(-1). These results point out to a fine-tuning mechanism for improving protein stability. Circular dichroism spectroscopic analysis of the unfolded state shows that most of the secondary and tertiary structures were disrupted. The fraction of Trp fluoresce...Continue Reading

References

Oct 1, 1995·Protein Science : a Publication of the Protein Society·J K MyersJ M Scholtz
Nov 1, 1995·Protein Science : a Publication of the Protein Society·C N PaceT Gray
May 13, 1997·Biochemistry·F W Lau, J U Bowie
Aug 10, 2001·Current Opinion in Structural Biology·J U Bowie
May 30, 2002·Biophysical Chemistry·John A Schellman
Dec 14, 2002·Biochimica Et Biophysica Acta·Valeria Levi, F Luis González Flecha
Sep 2, 2003·Biochimica Et Biophysica Acta·Michalis AivaliotisGeorgios Tsiotis
Sep 25, 2003·Proceedings of the National Academy of Sciences of the United States of America·Srebrenka RobicSusan Marqusee
Dec 9, 2003·Journal of Molecular Biology·Salem FahamJames U Bowie
May 14, 2004·Annual Review of Biophysics and Biomolecular Structure·Charles R Sanders, Jeffrey K Myers
Mar 19, 2005·The Journal of Biological Chemistry·Jean-Louis BanèresJoel Bockaert
Jul 20, 2005·Biochimica Et Biophysica Acta·Sharon M KellyNicholas C Price
Dec 31, 2005·Nucleic Acids Research·M D Shaji KumarAkinori Sarai
Jan 31, 2006·Biochimica Et Biophysica Acta·Bjarni Asgeirsson, Katrín Guojónsdóttir
Feb 24, 2006·The Journal of Biological Chemistry·Matthew H SazinskyAmy C Rosenzweig
Apr 8, 2006·Protein Science : a Publication of the Protein Society·Pankaj Sehgal, Daniel E Otzen
Jul 4, 2006·Current Opinion in Structural Biology·Paula J Booth, Paul Curnow
Jul 4, 2006·Protein Science : a Publication of the Protein Society·Abbas Razvi, J Martin Scholtz
Aug 16, 2006·Biochemistry·Matthew H SazinskyAmy C Rosenzweig
Oct 27, 2006·Biochimica Et Biophysica Acta·Ying YangJosé M Argüello
Jan 16, 2007·Biometals : an International Journal on the Role of Metal Ions in Biology, Biochemistry, and Medicine·José M ArgüelloManuel González-Guerrero
Jul 7, 2007·Physiological Reviews·Svetlana LutsenkoOleg Y Dmitriev
Jan 12, 2008·Archives of Biochemistry and Biophysics·Diego I CattoniJosé M Argüello
Apr 1, 2008·Journal of Molecular Biology·Sivaram ChintalapatiWerner Kühlbrandt
Apr 18, 2008·Proceedings of the National Academy of Sciences of the United States of America·Manuel González-Guerrero, José M Argüello
Sep 6, 2008·The Journal of Biological Chemistry·Manuel González-GuerreroJosé M Argüello
Jan 15, 2009·Proceedings of the National Academy of Sciences of the United States of America·Paul Curnow, Paula J Booth
Jan 23, 2009·Current Opinion in Structural Biology·Paula J Booth, Paul Curnow
Feb 5, 2009·Photochemistry and Photobiology·Craig McKibbinPaula J Booth
May 23, 2009·The FEBS Journal·Guang-Jun HeYong-Bin Yan
Aug 18, 2009·Biochimica Et Biophysica Acta·Diego I CattoniF Luis González Flecha

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Citations

Jun 23, 2012·PloS One·Martín M Dodes TraianF Luis González Flecha
Jun 28, 2014·Biomolecules·Ernesto A Roman, F Luis González Flecha
May 14, 2011·Biotechnology Journal·Bruno MantaAna Denicola
Mar 31, 2016·Biophysical Journal·Rashmi PanigrahiM Joanne Lemieux
Feb 18, 2014·Journal of Molecular Biology·Nicola J HarrisPaula J Booth
Dec 18, 2013·Biochimica Et Biophysica Acta·Oznur TastanJudith Klein-Seetharaman
Apr 19, 2016·Biochimica Et Biophysica Acta·Luis M Bredeston, F Luis González Flecha
Mar 15, 2011·Biochimica Et Biophysica Acta·Daniel Otzen
Aug 10, 2010·Biochimica Et Biophysica Acta·Iona P Garber CohenF Luis González Flecha
Nov 22, 2011·Biochimica Et Biophysica Acta·Nicola J Harris, Paula J Booth
Sep 19, 2017·Biophysics Reviews·F Luis González Flecha
Dec 13, 2012·Physical Chemistry Chemical Physics : PCCP·Bijan Kumar PaulNikhil Guchhait
May 9, 2014·Langmuir : the ACS Journal of Surfaces and Colloids·Bijan K PaulSaptarshi Mukherjee

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