title: Substrate profiling of Zika virus NS2B-NS3 protease.
abstract: Zika virus (ZIKV), isolated from macaques in Uganda in 1947, was not considered to be a dangerous human pathogen. However, this view has recently changed as ZIKV infections are now associated with serious pathological disorders including microcephaly and Guillain-Barré syndrome. Similar to other viruses in the Flaviviridae family, ZIKV expresses the serine protease NS3 which is responsible for viral protein processing and replication. Herein, we report the expression of an active NS3prodomain fused with the NS2B cofactor (NS2BLNNS3pro) in a prokaryotic expression system and profile its specificity for synthesized FRET-type substrate libraries. Our findings pave way for screening potential intracellular substrates of NS3 and for developing specific inhibitors of this ZIKV protease.
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Gdansk, Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Gruba
authors.affiliation: Department of Microbiology, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Jose Ignacio
authors.initials: JI
authors.lastname: Rodriguez Martinez
authors.affiliation: Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland.
authors.forename: Renata
authors.initials: R
authors.lastname: Grzywa
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Gdansk, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Wysocka
authors.affiliation: Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Skoreński
authors.affiliation: Department of Microbiology, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.|Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Burmistrz
authors.affiliation: Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland.
authors.forename: Maria
authors.initials: M
authors.lastname: Łęcka
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, University of Gdansk, Poland. adam.lesner@ug.edu.pl.
authors.forename: Adam
authors.initials: A
authors.lastname: Lesner
authors.affiliation: Division of Medicinal Chemistry and Microbiology, Faculty of Chemistry, Wroclaw University of Science and Technology, Wroclaw, Poland. marcin.sienczyk@pwr.edu.pl.
authors.forename: Marcin
authors.initials: M
authors.lastname: Sieńczyk
authors.affiliation: Department of Microbiology, Faculty of Biochemistry Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland. k.a.pyrc@uj.edu.pl.|Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland. k.a.pyrc@uj.edu.pl.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Pyrć
mesh_terms: D001665:Binding Sites; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D006801:Humans; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D019151:Peptide Library; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000378:metabolism*; D013379:Substrate Specificity; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000378:metabolism*; D000071244:Zika Virus / Q000737:chemistry / Q000201:enzymology*
chemical_list: D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins; D012697:Serine Endopeptidases
keywords: NS2B-NS3 protease; Zika virus; combinatorial chemistry; substrate library; substrate mapping