title: Exploring the activity of tobacco etch virus protease in detergent solutions.
abstract: Tobacco etch virus (TEV) protease is generally used to remove affinity tags from target proteins. It has been reported that some detergents inhibit the activity of this protease, and therefore should be avoided when removing affinity tags from membrane proteins. The aim of this study was to explore and evaluate this further. Hence, affinity tag removal with TEV protease was tested from three membrane proteins (a Pgp synthase and two CorA homologs) in the presence of 16 different detergents commonly used in membrane protein purification and crystallization. We observed that in the presence of the same detergent (Triton X-100), TEV protease could remove the affinity tag completely from one protein (CorA) but not from another protein (Pgp synthase). There was also a large variation in yield of cleaved membrane protein in different detergents, which probably depends on features of the protein-detergent complex. These observations show that, contrary to an earlier report, detergents do not inhibit the enzymatic activity of the TEV protease.
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Anna-Karin
authors.initials: AK
authors.lastname: Lundbäck
authors.forename: Susanne
authors.initials: S
authors.lastname: van den Berg
authors.forename: Hans
authors.initials: H
authors.lastname: Hebert
authors.forename: Helena
authors.initials: H
authors.lastname: Berglund
authors.forename: Said
authors.initials: S
authors.lastname: Eshaghi
mesh_terms: D003902:Detergents / Q000494:pharmacology*; D010450:Endopeptidases / Q000378:metabolism*; D017800:Potyvirus / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology*; D012996:Solutions
chemical_list: D003902:Detergents; D011480:Protease Inhibitors; D012996:Solutions; D010450:Endopeptidases; C099488:TEV protease