Wieligmann, Karin and Norledge, Brian and Jaenicke, Rainer and Mayr, Eva-Maria (1998) Eye lens beta B2-crystallin: Circular permutation does not influence the oligomerization state but enhances the conformational stability. JOURNAL OF MOLECULAR BIOLOGY, 280 (4). pp. 721-729. ISSN 0022-2836,
Full text not available from this repository.Abstract
The related vertebrate eye lens polypeptides, beta B2- and gamma B-crystallin, each fold into two similar beta-sheet domains. The main difference is the state of oligomerization resulting from intermolecular domain interactions in the oligomeric beta-crystallins and intramolecular contacts in the monomeric gamma-crystallins. The question arises whether it is possible to create a monomeric gamma B-like beta B2-molecule by protein engineering methods. We wanted to produce such a molecule by circularly permuting the domains of beta B2-crystallin. The new termini were created from the original connecting peptide, and the new linker from stumps of the original extensions, while the rest of the flexible extensions were deleted. As judged by circular dichroism and fluorescence, the permutation causes little change in the structure of the protein. The circularly permuted protein forms dimers as wild-type beta B2-crystallin. On the other hand, cp beta B2 shows a slightly enhanced stability against urea with a midpoint of transition of 2.1 M urea versus 1.9 M for the wild-type protein lacking N and C-terminal arms, thus indicating stronger domain interactions. To our knowledge this is the first circularly permuted protein which exhibits a higher stability than the corresponding wild-type protein. (C) 1998 Academic Press.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | C-TERMINAL EXTENSIONS; X-RAY-ANALYSIS; BETA-CRYSTALLINS; DIHYDROFOLATE-REDUCTASE; SECONDARY STRUCTURE; POLYPEPTIDE-CHAINS; PERMUTED VARIANTS; ESCHERICHIA-COLI; RNA-POLYMERASE; CLONED GENES; crystallins; circular permutation; domain association; oligomerization; protein stability |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 02 May 2023 13:29 |
| Last Modified: | 02 May 2023 13:29 |
| URI: | https://pred.uni-regensburg.de/id/eprint/49663 |
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