Faraone-Mennella, Jasmin and Tezcan, F. Akif and Gray, Harry B. and Winkler, Jay R. (2006) Stability and Folding Kinetics of Structurally Characterized Cytochrome c-b_(562). Biochemistry, 45 (35). pp. 10504-10511. ISSN 0006-2960. doi:10.1021/bi060242x. https://resolver.caltech.edu/CaltechAUTHORS:20170131-102250205
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Abstract
The four-helix-bundle protein fold can be constructed from a wide variety of primary amino acid sequences. Proteins with this structure are excellent candidates for investigations of the relationship between folding mechanism and topology. The folding of cytochrome b_(562), a four-helix-bundle heme protein, is hampered by heme dissociation. To overcome this complication, we have engineered a variant of cytochrome b_(562) (cyt c-b_(562)) featuring a c-type linkage between the heme and the polypeptide chain. The replacement of the native cyt b_(562) leader sequence in this protein with that of a c-type cytochrome (cyt c_(556)) led to high yields of fully matured and correctly folded cyt c-b_(562). We have determined the X-ray crystal structure of cyt c-b_(562) at 2.25 Å and characterized its physical, chemical, and folding properties. These measurements reveal that the c-type linkage does not perturb the protein fold or reduction potential of the heme group. The covalent attachment of the porphyrin to the polypeptide does, however, produce a substantial change in protein stability and folding kinetics.
| Item Type: | Article | |||||||||
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| Additional Information: | © 2006 American Chemical Society. Received 5 February 2006. Published online 10 August 2006. Published in print 1 September 2006. This work was supported by National Institutes of Health Grant GM068461 (J.R.W.) and the Ellison Medical Foundation (Senior Scholar Award in Aging to H.B.G.). | |||||||||
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| Issue or Number: | 35 | |||||||||
| DOI: | 10.1021/bi060242x | |||||||||
| Record Number: | CaltechAUTHORS:20170131-102250205 | |||||||||
| Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20170131-102250205 | |||||||||
| Official Citation: | Stability and Folding Kinetics of Structurally Characterized Cytochrome c-b562 Jasmin Faraone-Mennella, F. Akif Tezcan, Harry B. Gray, and Jay R. Winkler Biochemistry 2006 45 (35), 10504-10511 DOI: 10.1021/bi060242x | |||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||
| ID Code: | 73871 | |||||||||
| Collection: | CaltechAUTHORS | |||||||||
| Deposited By: | Ruth Sustaita | |||||||||
| Deposited On: | 31 Jan 2017 19:50 | |||||||||
| Last Modified: | 11 Nov 2021 05:23 |
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