title: Selection of suitable detergents for obtaining an active dengue protease in its natural form from E. coli.
abstract: Dengue protease is a two-component enzyme and is an important drug target against dengue virus. The protease activity and protein stability of dengue nonstructural protein 3 (NS3) require a co-factor region from a four-span membrane protein NS2B. A natural form of dengue protease containing full-length NS2B and NS3 protease domain NS2BFL-NS3pro will be useful for dengue drug discovery. In current study, detergents that can be used for protease purification were tested. Using a water soluble protease construct, 39 detergents were selected for both NS2B and NS2BFL-NS3pro purification. The results showed that 18 detergents were able to sustain the activity of the natural dengue protease and 11 detergents could be used for NS2B purification. The results obtained in this study will be useful for biochemical and biophysical studies on dengue protease.
authors: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31Biopolis Way, Nanos, #03-01, 138669, Singapore.
authors: Lynette Sin Yee
authors: LS
authors: Liew
authors: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31Biopolis Way, Nanos, #03-01, 138669, Singapore.
authors: Michelle Yueqi
authors: MY
authors: Lee
authors: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31Biopolis Way, Nanos, #03-01, 138669, Singapore.
authors: Ying Lei
authors: YL
authors: Wong
authors: Institute of Materials Research and Engineering, 3 Research Link, 117602, Singapore.
authors: Jinting
authors: J
authors: Cheng
authors: Institute of Chemical & Engineering Sciences, Agency for Science, Technology and Research (A*STAR), 1Pesek Road, Jurong Island, Singapore, 627833, Singapore. Electronic address: li_qingxin@ices.a-star.edu.sg.
authors: Qingxin
authors: Q
authors: Li
authors: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A*STAR), 31Biopolis Way, Nanos, #03-01, 138669, Singapore. Electronic address: cbkang@etc.a-star.edu.sg.
authors: CongBao
authors: C
authors: Kang
mesh_terms: D003715:Dengue / Q000201:enzymology* / Q000821:virology; D003716:Dengue Virus / Q000201:enzymology* / Q000472:pathogenicity; D003902:Detergents / Q000737:chemistry*; D055808:Drug Discovery; D004926:Escherichia coli / Q000235:genetics; D007700:Kinetics; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000302:isolation & purification; D000085242:Viral Proteases
chemical_list: D003902:Detergents; D012697:Serine Endopeptidases; D017361:Viral Nonstructural Proteins; C504449:NS3 protease, dengue virus; D000085242:Viral Proteases
keywords: Dengue protease; Dengue virus; Detergent micelles; Membrane protein