FIGURE 7.
Rapid reoxidation of DTT-reduced 147CC514. A, effect of aerobic preincubation on activity of the mutant protein with P450. B, effect of aerobic incubation of disulfide cross-linked DTT and both DTT- and iodoacetamide-treated mutant CYPOR on the ability of the protein to reduce cyt c. Note the rapid loss of activity of the DTT-treated protein. The data suggest iodoacetamide treatment does not alkylate all free thiols. The reactions were conducted as described under “Experimental Procedures.”