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Solution Structure of Oxidized Saccharomyces cerevisiae Iso-1-cytochrome c

Banci, Lucia and Bertini, Ivano and Bren, Kara L. and Gray, Harry B. and Sompornpisut, Pornthep and Turano, Paola (1997) Solution Structure of Oxidized Saccharomyces cerevisiae Iso-1-cytochrome c. Biochemistry, 36 (29). pp. 8992-9001. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20180213-072130274

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Abstract

The solution structure of oxidized Saccharomyces cerevisiae Cys102Ser iso-1-cytochrome c has been determined using 1361 meaningful NOEs (of 1676 total) after extending the published proton assignment [Gao, Y., et al. (1990) Biochemistry 29, 6994−7003] to 77% of all proton resonances. The NOE patterns indicate that secondary structure elements are maintained upon oxidation in solution with respect to the solid state and solution structures of the reduced species. Constraints derived from the pseudocontact shifts [diamagnetic reference shift values are those of the reduced protein [Baistrocchi, P., et al. (1996) Biochemistry 35, 13788−13796]] were used in the final stages of structure calculations. After restrained energy minimization with constraints from NOEs and pseudocontact shifts, a family of 20 structures with rmsd values of 0.58 ± 0.08 and 1.05 ± 0.10 Å (relative to the average structure) for the backbone and all heavy atoms, respectively, was obtained. The solution structure is compared with the crystal structure and the structures of related systems. Twenty-six amide protons were detected in the NMR spectrum 6 days after the oxidized lyophilized protein was dissolved in D_2O (pH 7.0 and 303 K); in an analogous experiment, 47 protons were observed in the spectrum of the reduced protein. The decrease in the number of nonexchanging amide protons, which mainly are found in the loop regions 14−26 and 75−82, confirms the greater flexibility of the structure of oxidized cytochrome c in solution. Our finding of increased solvent accessibility in these loop regions is consistent with proposals that an early step in unfolding the oxidized protein is the opening of the 70−85 loop coupled with dissociation of the Met80−iron bond.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bi963025cDOIArticle
https://pubs.acs.org/doi/full/10.1021/bi963025cPublisherArticle
https://pubs.acs.org/doi/suppl/10.1021/bi963025cPublisherSupporting Information
ORCID:
AuthorORCID
Gray, Harry B.0000-0002-7937-7876
Turano, Paola0000-0002-7683-8614
Additional Information:© 1997 American Chemical Society. Received 10 December 1996. Published online 22 July 1997. This work was supported by EC Biotechnology Program BIO2- CT94-2052 (DG 12 SSMA), the Italian CNR, and the United States NSF. Experiments were performed with the instrumentation of the Florence Laboratory of Relaxometry and Magnetic Resonance on Paramagnetic Metalloproteins, Large Scale Facility of the European Union.
Funders:
Funding AgencyGrant Number
European CommissionBIO2-CT94-2052 (DG 12 SSMA)
Consiglio Nazionale delle Ricerche (CNR)UNSPECIFIED
NSFUNSPECIFIED
Issue or Number:29
Record Number:CaltechAUTHORS:20180213-072130274
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20180213-072130274
Official Citation:Solution Structure of Oxidized Saccharomyces cerevisiae Iso-1-cytochrome c, Lucia Banci, Ivano Bertini, Kara L. Bren, Harry B. Gray, Pornthep Sompornpisut, and Paola Turano Biochemistry 1997 36 (29), 8992-9001 DOI: 10.1021/bi963025c
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84800
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:13 Feb 2018 21:01
Last Modified:22 Nov 2019 09:58

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