title: West Nile virus protease activity in detergent solutions and application for affinity tag removal.
abstract: Affinity tags are typically used to facilitate membrane protein purification. A protease needs to remain active in the presence of detergent to remove a fusion tag from a recombinant membrane protein. Our results show that the West Nile virus (WNV) protease activity was not affected while in the presence of a wide range of detergents. In a detergent solution, the WNV protease can remove the fusion tag from a recombinant protein containing KCNE3 and a WNV protease site. Therefore, the WNV protease may be useful as an alternative enzyme to remove affinity tags in protein purifications.
authors.affiliation: Experimental Therapeutics Centre, Agency for Science, Technology and Research (A∗STAR), Singapore 138669, Singapore.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Huang
authors.forename: Qingxin
authors.initials: Q
authors.lastname: Li
authors.forename: Angela Shuyi
authors.initials: AS
authors.lastname: Chen
authors.forename: Congbao
authors.initials: C
authors.lastname: Kang
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D003902:Detergents / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D024642:Potassium Channels, Voltage-Gated / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012996:Solutions; D014902:West Nile virus / Q000201:enzymology*
chemical_list: D003902:Detergents; C418492:KCNE3 protein, human; D010455:Peptides; D024642:Potassium Channels, Voltage-Gated; D011994:Recombinant Proteins; D012996:Solutions; D010447:Peptide Hydrolases