Structural insights on the efficient catalysis of hydroperoxide reduction by Ohr: Crystallographic and molecular dynamics approaches

Show simple item record Piccirillo, Erika Alegria, Thiago G. P. Discola, Karen F. Cussiol, Jose Renato Rosa [UNIFESP] Domingos, Renato M. Oliveira, Marcos A. de Rezende, Leandro de Netto, Luis E. S. Amaral, Antonia T-do 2018-07-26T12:18:46Z 2018-07-26T12:18:46Z 2018
dc.identifier.citation Plos One. San Francisco, v. 13, n. 5, p. -, 2018.
dc.identifier.issn 1932-6203
dc.description.abstract Organic hydroperoxide resistance (Ohr) enzymes are highly efficient Cys-based peroxidases that play central roles in bacterial response to fatty acid hydroperoxides and peroxynitrite, two oxidants that are generated during host-pathogen interactions. In the active site of Ohr proteins, the conserved Arg (Arg19 in Ohr from Xylella fastidiosa) and Glu (Glu51 in Ohr from Xylella fastidiosa) residues, among other factors, are involved in the extremely high reactivity of the peroxidatic Cys (C-p) toward hydroperoxides. In the closed state, the thiolate of C-p is in close proximity to the guanidinium group of Arg19. Ohr enzymes can also assume an open state, where the loop containing the catalytic Arg is far away from C-p and Glu51. Here, we aimed to gain insights into the putative structural switches of the Ohr catalytic cycle. First, we describe the crystal structure of Ohr from Xylella fastidiosa (XfOhr) in the open state that, together with the previously described XfOhr structure in the closed state, may represent two snapshots along the coordinate of the enzyme-catalyzed reaction. These two structures were used for the experimental validation of molecular dynamics (MD) simulations. MD simulations employing distinct protonation states and in silico mutagenesis indicated that the polar interactions of Arg19 with Glu51 and C-p contributed to the stabilization of XfOhr in the closed state. Indeed, C-p oxidation to the disulfide state facilitated the switching of the Arg19 loop from the closed to the open state. In addition to the Arg19 loop, other portions of XfOhr displayed high mobility, such as a loop rich in Gly residues. In summary, we obtained a high correlation between crystallographic data, MD simulations and biochemical/enzymatic assays. The dynamics of the Ohr enzymes are unique among the Cys-based peroxidases, in which the active site Arg undergoes structural switches throughout the catalytic cycle, while C-p remains relatively static. en
dc.description.sponsorship Fundacao de Amparo a Pesquisa no Estado de Sao Paulo (FAPESP), Brazil
dc.description.sponsorship Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), Brazil
dc.description.sponsorship Redox Processes in Biomedicine
dc.format.extent -
dc.language.iso eng
dc.publisher Public Library Science
dc.rights Acesso aberto
dc.title Structural insights on the efficient catalysis of hydroperoxide reduction by Ohr: Crystallographic and molecular dynamics approaches en
dc.type Artigo
dc.description.affiliation Univ Sao Paulo, Inst Quim, Dept Quim Fundamental, Sao Paulo, SP, Brazil
dc.description.affiliation Univ Sao Paulo, Inst Biociencias, Dept Genet & Biol Evolut, Sao Paulo, SP, Brazil
dc.description.affiliation Univ Estadual Paulista, Inst Biociencias, Campus Litoral Paulista, Sao Vicente, SP, Brazil
dc.description.affiliation Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, Brazil
dc.description.affiliationUnifesp Univ Fed Sao Paulo, Dept Bioquim, Sao Paulo, SP, Brazil
dc.description.sponsorshipID Redox: 13/07937-8
dc.description.sponsorshipID FAPESP: 2008/07971-3
dc.description.sponsorshipID FAPESP: 2009/12885-1
dc.description.sponsorshipID FAPESP: 2005/50056-6
dc.description.sponsorshipID FAPESP: 2014/01614-5
dc.description.sponsorshipID FAPESP: 2012/06633-2
dc.description.sponsorshipID FAPESP: 2016/12392-9
dc.identifier.file WOS000432537100011.pdf
dc.identifier.doi 10.1371/journal.pone.0196918
dc.description.source Web of Science
dc.identifier.wos WOS:000432537100011


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