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Enhancing the Thermal Stability of Avidin
In this study we showed that tetrameric chicken avidin can be stabilized by introducing intermonomeric disulfide bridges between its subunits. These covalent bonds had no major effects on the biotin binding properties of the respective mutants. Moreover, one of the mutants (Avd-ccci) maintained its...
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Published in: | The Journal of biological chemistry 2003-01, Vol.278 (4), p.2479-2483 |
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Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | In this study we showed that tetrameric chicken avidin can be stabilized by introducing intermonomeric disulfide bridges between
its subunits. These covalent bonds had no major effects on the biotin binding properties of the respective mutants. Moreover,
one of the mutants (Avd-ccci) maintained its tetrameric integrity even in denaturing conditions. The new avidin forms Avd-ci
and Avd-ccci, which have nativeâââdenatured transition midpoints ( T m ) of 98.6 and 94.7â°C, respectively, in the absence of biotin, will find use in applications where extreme stability or minimal
leakage of subunits is required. Furthermore, we showed that the intramonomeric disulfide bridges found in the wild-type avidin
affect its stability. The mutant Avd-nc, in which this bridge was removed, had a lower T m in the absence of biotin than the wild-type avidin but showed comparable stability in the presence of biotin. |
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ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M210721200 |