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

Debora N. Okamoto, Lilian C.G. Oliveira, Marcia Y. Kondo, Maria H.S. Cezari, Zoltán Szeltner, Tünde Juhász, Maria A. Juliano, László Polgár, Luiz Juliano, Iuri E. Gouvea

Research output: Contribution to journalArticle

9 Citations (Scopus)

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 Na2SO4, 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 (Kd) 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 kcat increase and Km 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.

Original languageEnglish
Pages (from-to)1461-1468
Number of pages8
JournalBiological Chemistry
Volume391
Issue number12
DOIs
Publication statusPublished - Dec 1 2010

Keywords

  • Coronavirus
  • Hofmeister series
  • Kosmotropic salt
  • Macromolecular crowding
  • Molecular crowding
  • Protease

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Clinical Biochemistry

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  • Cite this

    Okamoto, D. N., Oliveira, L. C. G., Kondo, M. Y., Cezari, M. H. S., Szeltner, Z., Juhász, T., Juliano, M. A., Polgár, L., Juliano, L., & Gouvea, I. E. (2010). Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition. Biological Chemistry, 391(12), 1461-1468. https://doi.org/10.1515/BC.2010.145