Alteration of human myoglobin proximal histidine to cysteine or tyrosine by site-directed mutagenesis: characterization and their catalytic activities.

Two mutant proteins of human myoglobin (Mb) that exhibit altered axial ligations were prepared by site-directed mutagenesis of a cloned gene for human Mb. The normal axial ligand residue, histidine 93(F8), was replaced with cysteine or tyrosine, resulting in H93C or H93Y Mb, respectively. Cysteine or tyrosine coordination to the ferric heme iron is verified by electronic absorption, 1H-NMR, EPR spectra, and redox potentials of Fe2+/Fe3+ couple. Their mono-oxygenation activities of styrene are also discussed.

[1]  Carl Djerassi,et al.  Magnetic circular dichroism studies. 43. Oxidized cytochrome P-450. Magnetic circular dichroism evidence for thiolate ligation in the substrate-bound form. Implications for the catalytic mechanism , 1976 .

[2]  N. Xuong,et al.  The crystal structure of cytochrome c peroxidase. , 1980, The Journal of biological chemistry.

[3]  R. Frankel,et al.  Axial ligation modes in iron(III) Porphyrins. Models for the oxidized reaction states of cytochrome P-450 enzymes and the molecular structure of iron(III) protoporphyrin IX dimethyl ester p-nitrobenzenethiolate. , 1976, Journal of the American Chemical Society.

[4]  M. Nagai,et al.  Reduction of methemoglobins M Hyde Park, M Saskatoon, and M Milwaukee by ferredoxin and ferredoxin-nicotinamide adenine dinucleotide phosphate reductase system. , 1983, The Journal of biological chemistry.

[5]  S. Boxer,et al.  Electrostatic interactions in wild-type and mutant recombinant human myoglobins. , 1989, Biochemistry.

[6]  T. Inubushi,et al.  Nuclear magnetic resonance studies of high-spin ferric hemoproteins. , 1978, Advances in biophysics.

[7]  H. Yamada,et al.  Effects of 2,4-substituents of deuteropheme upon redox potentials of horseradish peroxidases. , 1975, Archives of biochemistry and biophysics.

[8]  P. Ortiz de Montellano,et al.  Epoxidation of styrene by hemoglobin and myoglobin. Transfer of oxidizing equivalents to the protein surface. , 1985, The Journal of biological chemistry.

[9]  J. Dawson,et al.  Cytochrome P-450 and chloroperoxidase: thiolate-ligated heme enzymes. Spectroscopic determination of their active-site structures and mechanistic implications of thiolate ligation , 1987 .

[10]  I. C. Gunsalus,et al.  Cytochrome P-450cam. I. Crystallization and properties. , 1974, The Journal of biological chemistry.

[11]  B C Finzel,et al.  The 2.6-A crystal structure of Pseudomonas putida cytochrome P-450. , 1985, The Journal of biological chemistry.

[12]  T. Kitagawa,et al.  Characteristics in tyrosine coordinations of four hemoglobins M probed by resonance Raman spectroscopy. , 1989, Biochemistry.

[13]  T. J. Reid,et al.  Structure of beef liver catalase. , 1981, Journal of molecular biology.

[14]  S. Martinis,et al.  Alteration of sperm whale myoglobin heme axial ligation by site-directed mutagenesis. , 1990, Biochemistry.

[15]  H. Morimoto,et al.  Properties of hemoglobin M, Milwaukee-I variant and its unique characteristic. , 1969, Biochimica et biophysica acta.

[16]  Y. Ogura,et al.  Studies on the EPR and light absorption spectra of horse erythrocyte catalase and its derivatives. , 1968, Journal of biochemistry.