Inhibitor and substrate binding induced stability of HIV-1 protease against sequential dissociation and unfolding revealed by high pressure spectroscopy and kinetics

PloS One
Marek IngrEliane Dumay

Abstract

High-pressure methods have become an interesting tool of investigation of structural stability of proteins. They are used to study protein unfolding, but dissociation of oligomeric proteins can be addressed this way, too. HIV-1 protease, although an interesting object of biophysical experiments, has not been studied at high pressure yet. In this study HIV-1 protease is investigated by high pressure (up to 600 MPa) fluorescence spectroscopy of either the inherent tryptophan residues or external 8-anilino-1-naphtalenesulfonic acid at 25°C. A fast concentration-dependent structural transition is detected that corresponds to the dimer-monomer equilibrium. This transition is followed by a slow concentration independent transition that can be assigned to the monomer unfolding. In the presence of a tight-binding inhibitor none of these transitions are observed, which confirms the stabilizing effect of inhibitor. High-pressure enzyme kinetics (up to 350 MPa) also reveals the stabilizing effect of substrate. Unfolding of the protease can thus proceed only from the monomeric state after dimer dissociation and is unfavourable at atmospheric pressure. Dimer-destabilizing effect of high pressure is caused by negative volume change of dimer ...Continue Reading

References

Sep 15, 1991·European Journal of Biochemistry·D Deville-Bonne, A J Else
Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·Y S ChengC A Kettner
May 3, 1988·Biochemistry·K Ruan, G Weber
Mar 31, 1993·Biochemical and Biophysical Research Communications·P Kuzmic
May 6, 1996·Biochemical and Biophysical Research Communications·T UhlíkováJ Vondrásek
Jan 8, 1999·Journal of Molecular Biology·B G Turner, M F Summers
Aug 19, 1999·Protein Science : a Publication of the Protein Society·D XieJ W Erickson
Mar 18, 2000·Biochemical and Biophysical Research Communications·K RuanC Balny
Oct 25, 2000·Protein Science : a Publication of the Protein Society·K StrisovskyH G Kräusslich
Oct 9, 2001·Trends in Biochemical Sciences·J L SilvaC A Royer
Sep 23, 2003·Protein Science : a Publication of the Protein Society·Marek IngrJan Konvalinka
Oct 22, 2003·Journal of Molecular Biology·Václav VeverkaRichard Hrabal
Sep 18, 2004·European Journal of Biochemistry·M Judith KornblattClaude Balny
Aug 6, 2005·Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Médicas E Biológicas·S MarchalC Balny
Apr 7, 2006·Protein Science : a Publication of the Protein Society·Josep FontMaria Vilanova
Feb 27, 2007·Biochimica Et Biophysica Acta·Oktay K Gasymov, Ben J Glasgow
Apr 7, 2007·The Journal of Biological Chemistry·Rieko IshimaJohn M Louis
Oct 3, 2009·Protein Science : a Publication of the Protein Society·John M LouisJane M Sayer
Dec 1, 2009·Viruses·Jana PokornáJan Konvalinka
Dec 5, 2012·PloS One·Stephane MarchalSabato D'Auria
Mar 13, 2014·Viral Immunology·Daniel Ferreira de Lima NetoClarice Weis Arns
Jul 23, 2014·Journal of Proteomics & Bioinformatics·Carol B FowlerJeffrey T Mason

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Citations

Jul 12, 2018·Scientific Reports·Jana HumpolíčkováTaťána Majerová
May 14, 2016·Journal of Molecular Biology·John M Louis, Julien Roche

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Methods Mentioned

BETA
NMR

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