Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition

Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition

Author Okamoto, Debora N. Autor UNIFESP Google Scholar
Oliveira, Lilian C. G. Autor UNIFESP Google Scholar
Kondo, Marcia Y. Autor UNIFESP Google Scholar
Cezari, Maria H. S. Autor UNIFESP Google Scholar
Szeltner, Zoltan Google Scholar
Juhasz, Nde Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Polgar, Laszlo Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Gouvea, Iuri E. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Hungarian Acad Sci
Abstract The 3C-like peptidase of the severe acute respiratory syndrome virus (SARS-CoV) is strictly required for viral replication, thus being a potential target for the development of antiviral agents. in contrast to monomeric picornavirus 3C peptidases, SARS-CoV 3CLpro exists in equilibrium between the monomer and dimer forms in solution, and only the dimer is proteolytically active in dilute buffer solutions. in this study, the increase of SARS-CoV 3CLpro peptidase activity in presence of kosmotropic salts and crowding agents is described. the activation followed the Hofmeister series of anions, with two orders of magnitude enhancement in the presence of Na(2)SO(4), whereas the crowding agents polyethylene glycol and bovine serum albumin increased the hydrolytic rate up to 3 times. Kinetic determinations of the monomer dimer dissociation constant (K(d)) indicated that activation was a result of a more active dimer, without significant changes in K(d) values. the activation was found to be independent of substrate length and was derived from both k(cat) increase and K(m) decrease. the viral peptidase activation described here could be related to the crowded intracellular environment and indicates a further fine-tuning mechanism for biological control, particularly in the microenvironment of the vesicles that are induced in host cells during positive strand RNA virus infection.
Keywords coronavirus
Hofmeister series
kosmotropic salt
macromolecular crowding
molecular crowding
protease
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Date 2010-12-01
Published in Biological Chemistry. Berlin: Walter de Gruyter & Co, v. 391, n. 12, p. 1461-1468, 2010.
ISSN 1431-6730 (Sherpa/Romeo, impact factor)
Publisher Walter de Gruyter & Co
Extent 1461-1468
Origin http://dx.doi.org/10.1515/BC.2010.145
Access rights Closed access
Type Article
Web of Science ID WOS:000284586700011
URI http://repositorio.unifesp.br/handle/11600/33160

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