Enzymatic characterization of a trypsin-like serine protease encoded by the genome of Cell fusing agent virus
Autor(es): Peyrefitte, Christophe N; Pastorino, Boris A M; Grandadam, Marc; Rolland, Dominique; Tolou, Hugues J; Bessaud, Maël
Resumo: Cell fusing agent virus (CFAV) is a positive strand RNA insect virus first isolated from a mosquito cell line. Based on viral morphology, phenotypic and phylogenetic studies, CFAV had been tentatively assigned to the genus Flavivirus (family Flaviviridae ). The determination of the CFAV polyprotein complete sequence showed a putative serine protease domain analogue to the flaviviral NS2B/NS3 complex. This complex had been extensively studied, because it represented one of the main targets for antiflavivirus therapy development. We report herein the biochemical characterization of CFAV ?NS2B-NS3pro protease complex. CFAV polyprotein sequence was computationally analysed to identify the amino-acid regions involved in protease activity. We designed, expressed and purified a catalytically active protease whose enzymatic properties were determined using fluorogenic substrates. Our results showed that, despite the low level of conservation of its amino-acid sequence, CFAV protease exhibited physico-chemical properties of other flaviviruses (high pH value requirement for optimal activity, inhibition by salt and preference for substrates featuring a basic residue at P 1 position).
Palavras-Chave: Amino acid sequence, Molecular sequence data, Serine endopeptidases - Isolation & purification
Imprenta: Virus Genes, v. 34, n. 2, p. 185-191, 2007.
Descritores: Zika virus - Cell ; Zika virus - Flaviviridae ; Zika virus - Genome ; Zika virus - Molecular structure ; Zika virus - Proteome ; Zika virus - RNA ; Zika virus - Biochemistry ; Zika virus - Protein synthesis ; Zika virus - Proteins ; Zika virus - Antibodies ; Zika virus - Molecular methods ; Zika virus - RNA virus
Data de publicação: 2007
