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The importance of b...
The importance of being knotted : Effects of the C-terminal knot structure on enzymatic and mechanical properties of bovine carbonic anhydrase II1
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- Alam, M.T. (författare)
- Laboratory of Biodynamics, Grad. Sch. of Biosci. and Biotech., Tokyo Inst. of Technol., 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
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- Yamada, T. (författare)
- Laboratory of Biodynamics, Grad. Sch. of Biosci. and Biotech., Tokyo Inst. of Technol., 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
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- Carlsson, Uno (författare)
- Linköpings universitet,Tekniska högskolan,Biokemi
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- Ikai, A. (författare)
- Laboratory of Biodynamics, Grad. Sch. of Biosci. and Biotech., Tokyo Inst. of Technol., 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan
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Laboratory of Biodynamics, Grad Sch. of Biosci. and Biotech., Tokyo Inst. of Technol., 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan Tekniska högskolan (creator_code:org_t)
- 2002
- 2002
- Engelska.
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Ingår i: FEBS Letters. - 0014-5793 .- 1873-3468. ; 519:1-3, s. 35-40
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- In order to better understand the contribution of the knotted folding pattern to the enzymatic and mechanical properties of carbonic anhydrases, we replaced Gln-253 of bovine carbonic anhydrase II with Cys, which allowed us to measure the mechanical strength of the protein against tensile deformation by avoiding knot tightening. The expressed protein, to our surprise, turned out to contain two conformational isomers, one capable of binding an enzymatic inhibitor and the other not, which led to their separation through affinity chromatography. In near- and far-UV circular dichroism and fluorescence spectra, the separated conformers were very similar to each other and to the wild-type enzyme, indicating that they both had native-like conformations. We describe new evidence which supports the notion that the difference between the two conformers is likely to be related to the completeness of the C-terminal knot formation. © 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
Nyckelord
- Atomic force microscope
- Carbonic anhydrase II
- Conformer
- Knot structure
- Mechanical unfolding
- TECHNOLOGY
- TEKNIKVETENSKAP
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