Concentration of Cu, EPR-detectable Cu, and formation of cupric-ferrocyanide in membranes with pMMO1

Add time:08/30/2019    Source:sciencedirect.com

EPR spectra were obtained for the type 2 Cu(2+) site in particulate methane monooxygenase, pMMO, from membrane fractions of Methylomicrobium album BG8. In addition to the EPR signal with g‖=2.24 and A‖=185 G found in both cells and membrane fractions, a second EPR signal with g‖=2.29 and A‖=146 G was found in membrane fractions and attributed to oxidation of cuprous sites. Comparison of EPR-detectable Cu(2+) with total copper determined by atomic absorption suggests that there are two or three EPR-silent coppers for every EPR-detectable copper and that there are approximately four coppers per enzyme composed of the 47, 27, and 25 kDa subunits. Treatment of membrane fractions loaded with pMMO with Fe(CN)3−6 results in a new EPR signal that is attributed to CuFe(CN)2−6, not to an intrinsic trimeric copper cluster as previously reported in studies with a related bacterium.

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