Three-Dimensional Solution Structure of Saccharomyces cerevisiae Reduced Iso-1-cytochrome c

• Published in: Biochemistry (Easton) 1996-10, Vol.35 (43), p.13788-13796
• Main Authors: Baistrocchi, Paolo, Banci, Lucia, Bertini, Ivano, Turano, Paola, Bren, Kara L, Gray, Harry B
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Crystallography, X-Ray; Cytochrome c Group - chemistry; Cytochrome-c Peroxidase - metabolism; Cytochromes c; Hydrogen Bonding; Iron - metabolism; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Mutation - genetics; Oxidation-Reduction; Protein Conformation; Protein Structure, Secondary; Protein Structure, Tertiary; Saccharomyces cerevisiae; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae Proteins
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi961110e

Two-dimensional 1H NMR spectra of Saccharomyces cerevisiae reduced iso-1-cytochrome c have been used to confirm and slightly extend the assignment available in the literature. 1702 NOESY cross-peaks have been assigned, and their intensities have been measured. Through the program DIANA and related protocols (Güntert, 1992), a solution structure has been obtained by using 1442 meaningful NOEs and 13 hydrogen-bond constraints. The RMSD values with respect to the mean structure for the backbone and all heavy atoms for a family of 20 structures are 0.61 ± 0.09 and 0.98 ± 0.09 Å, the average target function value being as small as 0.57 Å2. The larger number of slowly exchanging amide NHs observed in this system compared to that observed in the cyanide derivative of oxidized Ala 80 cytochrome c suggests that the oxidized form is much more flexible and that the backbone protons are more solvent accessible. Comparison of the present structure with the crystal structures of reduced yeast cytochrome c and of the complex between cytochrome c peroxidase and oxidized yeast cytochrome c reveals substantial similarity among the backbone conformations but differences in the residues located in the region of protein−protein interaction. Interestingly, in solution the peripheral residues involved in the interaction with cytochrome c peroxidase are on average closer to the position found in the crystal structure of the complex than to the solid state structure of the isolated reduced form.