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Stability and Folding Kinetics of Structurally Characterized Cytochrome c-b_(562)

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.

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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
Related URLs:
URLURL TypeDescription
Tezcan, F. Akif0000-0002-4733-6500
Gray, Harry B.0000-0002-7937-7876
Winkler, Jay R.0000-0002-4453-9716
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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Ellison Medical FoundationUNSPECIFIED
Issue or Number:35
Record Number:CaltechAUTHORS:20170131-102250205
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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
Deposited By: Ruth Sustaita
Deposited On:31 Jan 2017 19:50
Last Modified:11 Nov 2021 05:23

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