Direct measurement of the self-exchange rate of stellacyanin by a novel e.p.r. method

A method for reconstituting the blue copper protein stellacyanin with the stable copper isotopes 63Cu and 65Cu is reported. Small differences in the e.p.r. spectra of the two isotopic forms of stellacyanin have been used to monitor the electron self-exchange reaction of stellacyanin by rapid-freeze e.p.r. methods. The self-exchange rate constant (k11) for stellacyanin has been determined as 1.2 × 10(5) M-1 X S-1 at 20 degrees C. This value is in close agreement with values obtained from less-direct methods.