Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition
-
Debora N. Okamoto
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 Kd 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.
©2010 by Walter de Gruyter Berlin New York
Articles in the same Issue
- Minireview
- Physiology and pathophysiology of the RANKL/RANK system
- Genes and Nucleic Acids
- Murine aldo-keto reductase family 1 subfamily B: identification of AKR1B8 as an ortholog of human AKR1B10
- Characterization of plant miRNAs and small RNAs derived from potato spindle tuber viroid (PSTVd) in infected tomato
- Protein Structure and Function
- Characterization of a mutant R11H αB-crystallin associated with human inherited cataract
- Secretion of hepatoma-derived growth factor is regulated by N-terminal processing
- Twin arginine translocation (Tat)-dependent protein transport: the passenger protein participates in the initial membrane binding step
- Kinetic and structural characterization of bacterial glutaminyl cyclases from Zymomonas mobilis and Myxococcus xanthus
- Membranes, Lipids, Glycobiology
- Quantitative determination of haptoglobin glycoform variants in psoriasis
- Molecular Medicine
- Bile acid retention and activation of endogenous hepatic farnesoid-X-receptor in the pathogenesis of fatty liver disease in ob/ob-mice
- Cell Biology and Signaling
- Acute and long-term effects of metformin on the function and insulin secretory responsiveness of clonal β-cells
- Proteolysis
- Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition
- Pharmacological and genetic evidence that cathepsin B is not the physiological activator of rodent prorenin
- Acknowledgement
- Acknowledgement
Articles in the same Issue
- Minireview
- Physiology and pathophysiology of the RANKL/RANK system
- Genes and Nucleic Acids
- Murine aldo-keto reductase family 1 subfamily B: identification of AKR1B8 as an ortholog of human AKR1B10
- Characterization of plant miRNAs and small RNAs derived from potato spindle tuber viroid (PSTVd) in infected tomato
- Protein Structure and Function
- Characterization of a mutant R11H αB-crystallin associated with human inherited cataract
- Secretion of hepatoma-derived growth factor is regulated by N-terminal processing
- Twin arginine translocation (Tat)-dependent protein transport: the passenger protein participates in the initial membrane binding step
- Kinetic and structural characterization of bacterial glutaminyl cyclases from Zymomonas mobilis and Myxococcus xanthus
- Membranes, Lipids, Glycobiology
- Quantitative determination of haptoglobin glycoform variants in psoriasis
- Molecular Medicine
- Bile acid retention and activation of endogenous hepatic farnesoid-X-receptor in the pathogenesis of fatty liver disease in ob/ob-mice
- Cell Biology and Signaling
- Acute and long-term effects of metformin on the function and insulin secretory responsiveness of clonal β-cells
- Proteolysis
- Increase of SARS-CoV 3CL peptidase activity due to macromolecular crowding effects in the milieu composition
- Pharmacological and genetic evidence that cathepsin B is not the physiological activator of rodent prorenin
- Acknowledgement
- Acknowledgement
