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The Conformational Flexibility of Oxidized Cytochrome c Studied through Its Interaction with NH 3 and at High Temperatures
Author(s) -
Banci Lucia,
Bertini Ivano,
Spyroulias Georgios A.,
Turano Paola
Publication year - 1998
Publication title -
european journal of inorganic chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.667
H-Index - 136
eISSN - 1099-0682
pISSN - 1434-1948
DOI - 10.1002/(sici)1099-0682(199805)1998:5<583::aid-ejic583>3.0.co;2-y
Subject(s) - chemistry , adduct , cyanide , cytochrome c , ligand (biochemistry) , cytochrome , heme , proton nmr , crystallography , electron paramagnetic resonance , stereochemistry , hemeprotein , inorganic chemistry , nuclear magnetic resonance , organic chemistry , biochemistry , physics , receptor , mitochondrion , enzyme
The binding of ammonia to oxidized horse heart cytochrome c has been studied by 1 H‐NMR, EPR, and CD spectroscopy at pH = 8.0. The affinity constant of the ligand is in the range 1.5−4 M −1 . The 1 H‐NMR spectra of the heme group have been found to be similar to those of the high‐pH forms, high‐temperature forms, and cyanide adduct of the Met80Ala mutant of S . cerevisiae iso‐1‐cytochrome c. The assignment of a number of signals has led to the determination of the values of the magnetic anisotropy and of the orientation of its axes. The latter are similar to those of the Met80Ala cyanide derivative. The assignment of the high‐temperature species has been further pursued during this research. The analysis of the NMR data of the NH 3 adduct leads to the conclusion that substitution of Met80 at high pH or high temperature occurs through a ligand with cylindrical symmetry. This supports the suggestion that Met80 is substituted by a lysine at high pH.