Jose A Villegas, A Grant Mauk, Rafael Vazquez-Duhalt

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A cytochrome c variant resistant to heme degradation by hydrogen peroxide Jose A Villegas, A Grant Mauk, Rafael Vazquez-Duhalt Chemistry & Biology Volume 7, Issue 4, Pages 237-244 (April 2000) DOI: 10.1016/S1074-5521(00)00098-3

Figure 1 Inactivation and heme destruction of iso-1-cytochrome c in the presence of 1 mM H2O2. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)

Figure 2 The effect of hydrogen peroxide concentration on the rate constant for inactivation of iso-1-cytochrome c. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)

Figure 3 Heme prosthetic group destruction as indicated by disappearance of the Soret band in the presence of 1 mM H2O2. Spectra were recorded every 2 minutes, and the arrow indicates the direction of change. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)

Figure 4 Amino acid composition of iso-1-cytochrome c before and after incubation for 8 minutes in 1 mM H2O2. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)

Figure 5 Absorbance decrease of the Soret band (408 nm) of cytochrome c variants in the presence of 1 mM H2O2. Y– is a protein devoid of tyrosine which was prepared by PCR with various amplifications steps. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)

Figure 6 Direct ESR spectra obtained from the reaction of the wild-type (WT) yeast cytochrome c or the mutant Ile52→Phe67 with hydrogen peroxide. The reaction was performed with 250 μM cytochrome c and 0.25 M hydrogen peroxide (1:1000 ratio) in 20 mM phosphate buffer pH 7.2. After the addition of the peroxide, the reaction mixture was frozen in liquid nitrogen (within 15 seconds). The spectra were recorded at 4° under the following parameters: frequency 9.45 GHz; microwave power 0.5 mW; receiver gain 1 × 105; modulation frequency 100 KHz; modulation amplitude 3.89 G . The inset shows the effect of increasing temperature on the signal intensity. Chemistry & Biology 2000 7, 237-244DOI: (10.1016/S1074-5521(00)00098-3)