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

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dc.title Inhibitor and substrate binding induced stability of HIV-1 protease against sequential dissociation and unfolding revealed by high pressure spectroscopy and kinetics en
dc.contributor.author Ingr, Marek
dc.contributor.author Lange, Reinhard
dc.contributor.author Halabalová, Věra
dc.contributor.author Yehya, Alaa
dc.contributor.author Hrnčiřík, Josef
dc.contributor.author Chevalier-Lucia, Dominique
dc.contributor.author Palmade, Laetitia
dc.contributor.author Blayo, Claire
dc.contributor.author Konvalinka, Jan
dc.contributor.author Dumay, Eliane
dc.relation.ispartof PLoS ONE
dc.identifier.issn 1932-6203 OCLC, Ulrich, Sherpa/RoMEO, JCR
dc.date.issued 2015
utb.relation.volume 10
utb.relation.issue 3
dc.type article
dc.language.iso en
dc.publisher Public Library of Science
dc.identifier.doi 10.1371/journal.pone.0119099
dc.relation.uri http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0119099
dc.description.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 dissociation of -32.5 mL/mol. It helps us to determine the atmospheric pressure dimerization constant of 0.92 μM. High-pressure methods thus enable the investigation of structural phenomena that are difficult or impossible to measure at atmospheric pressure. © 2015 Ingr et al. en
utb.faculty Faculty of Technology
dc.identifier.uri http://hdl.handle.net/10563/1004241
utb.identifier.obdid 43873386
utb.identifier.scopus 2-s2.0-84924975892
utb.identifier.wok 000351284600050
utb.identifier.coden POLNC
utb.source j-scopus
dc.date.accessioned 2015-05-22T08:01:31Z
dc.date.available 2015-05-22T08:01:31Z
dc.description.sponsorship INSERM; Grant Agency of the Czech Republic [P208-12-G016]
dc.rights Attribution-NonCommercial-NoDerivs 4.0 International
dc.rights.uri http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.access openAccess
utb.contributor.internalauthor Ingr, Marek
utb.contributor.internalauthor Halabalová, Věra
utb.contributor.internalauthor Hrnčiřík, Josef
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