Exploring the activity of tobacco etch virus protease in detergent solutions |
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| Authors: | Lundbäck Anna-Karin van den Berg Susanne Hebert Hans Berglund Helena Eshaghi Said |
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| Affiliation: | aDivision of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-17177 Stockholm, Sweden;bKarolinska Institute, Department of Biosciences and Nutrition and Royal Institute of Technology, School of Technology and Health, Novum, SE-141 57 Huddinge, Sweden;cStructural Genomics Consortium, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-17177 Stockholm, Sweden |
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| 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. |
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