Role of the aromatic ring of Tyr43 in tetraheme cytochrome c(3) from Desulfovibrio vulgaris Miyazaki F.
Tyrosine 43 is positioned parallel to the fifth heme axial ligand, His34, of heme 1 in the tetraheme cytochrome c(3). The replacement of tyrosine with leucine increased the redox potential of heme 1 by 44 and 35 mV at the first and last reduction steps, respectively; its effects on the other hemes are small. In contrast, the Y43F mutation hardly changed the potentials. It shows that the aromatic ring at this position contributes to lowering the redox potential of heme 1 locally, although this cannot be the major contribution to the extremely low redox potentials of cytochrome c(3). Furthermore, temperature-dependent line-width broadening in partially reduced samples established that the aromatic ring at position 43 participates in the control of the kinetics of intramolecular electron transfer. The rate of reduction of Y43L cytochrome c(3) by 5-deazariboflavin semiquinone under partially reduced conditions was significantly different from that of the wild type in the last stage of the reduction, supporting the involvement of Tyr43 in regulation of reduction kinetics. The mutation of Y43L, however, did not induce a significant change in the crystal structure.
Institute for Protein Research, Osaka University, Suita, Japan.
Kiyoshi
K
Ozawa
Yuki
Y
Takayama
Fumiko
F
Yasukawa
Tomoaki
T
Ohmura
Michael A
MA
Cusanovich
Yusuke
Y
Tomimoto
Hideaki
H
Ogata
Yoshiki
Y
Higuchi
Hideo
H
Akutsu
D019943:Amino Acid Substitution; D003198:Computer Simulation; D003574:Cytochrome c Group / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000528:radiation effects*; D016969:Desulfovibrio vulgaris / Q000737:chemistry* / Q000201:enzymology*; D004789:Enzyme Activation; D004795:Enzyme Stability; D006418:Heme / Q000737:chemistry*; D006841:Hydrocarbons, Aromatic / Q000737:chemistry; D007834:Lasers; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D010782:Photolysis; D011487:Protein Conformation; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D014443:Tyrosine / Q000737:chemistry*
D003574:Cytochrome c Group; D006841:Hydrocarbons, Aromatic; D011994:Recombinant Proteins; D014443:Tyrosine; D006418:Heme; C017165:cytochrome c(3)