Yellow fever virus NS2B/NS3 protease: Hydrolytic Properties and Substrate Specificity

Yellow fever virus NS2B/NS3 protease: Hydrolytic Properties and Substrate Specificity

Author Kondo, Marcia Yuri Autor UNIFESP Google Scholar
Oliveira, Lilian Caroline Gonçalves de Autor UNIFESP Google Scholar
Okamoto, Débora Noma Autor UNIFESP Google Scholar
Araujo, Marina R. T. de Google Scholar
Duarte dos Santos, Claudia N. Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Gouvea, Iuri Estrada Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Inst Carlos Chagas Fiocruz PR
Abstract Here we report the hydrolytic behavior of recombinant YFV NS2B/NS3 protease against FRET substrates mimicking the prime and non-prime region of the natural polyprotein cleavage sites. While the P2-P'1 motif is the main factor associated with the catalytic efficiency of Dengue (DV) and West Nile Virus (WNV) protease, we show that the k(cat)/K-m of YFV NS2B/NS3 varied by more than two orders of magnitude, despite the presence of the same motif in all natural substrates. the catalytic significance of this homogeneity - a unique feature among worldwide prominent flavivirus - was kinetically analyzed using FRET peptides containing all possible combinations of two and three basic amino acids in tandem, and Arg and Lys residues produced distinct effects on k(cat)/K-m. the parallel of our data with those obtained in vivo by Chambers et al. (1991) restrains the idea that these sites co-evolved with the NS2B/NS3 protease to promote highly efficient hydrolysis and supports the notion that secondary substrate interaction distant from cleavage sites are the main factor associated with the different hydrolytic rates on YFV NS2B-NS3pro natural substrates. (C) 2011 Elsevier Inc. All rights reserved.
Keywords FRET substrate
Enzyme kinetics
Serine-proteases
Flavivirus
Dengue
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Date 2011-04-22
Published in Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 407, n. 4, p. 640-644, 2011.
ISSN 0006-291X (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 640-644
Origin http://dx.doi.org/10.1016/j.bbrc.2011.03.054
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000290240500003
URI http://repositorio.unifesp.br/handle/11600/33639

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