Resonance Raman study of the aa3-type cytochrome oxidase of thermophilic bacterium PS3.
Resonance Raman spectra of the aa3-type cytochrome oxidase of thermophilic bacterium PS3, which has a simpler subunit composition than the mitochondrial enzymes but very similar enzymatic properties, are investigated under various conditions and compared with those of mitochondrial enzymes. The intensities of the two marker lines of reduced cytochrome a3 at 1667 and 213 cm-1 had different dependences on the incubation temperatures and pH. With regard to the incubation temperature dependence, the intensity of the 1667-cm-1 line, the peripheral CH = O stretching mode of the a3 heme, behaved in nearly the same way as that of the oxidase activity whereas the intensity of the 213-cm-1 line, the Fe-histidine stretching mode of the a3 heme, exhibited a similar dependence to that of the proton pumping activity. The 213-cm-1 line disappeared upon binding of carbon monoxide, upon raising the pH above 9.2, or after incubating above 55 degrees C. The Raman line at 1611 cm-1, which was recently suggested to probe the proton pump activity [Babcock, G.T., & Callahan, P.M. (1983) Biochemistry 22, 2314-2319], remained unaltered after incubation at 60 degrees C for 20 min despite a reduction of proton pumping activity to one-third. This argues against the proposed mechanism. The frequencies of the Raman lines were the same for the intact membrane and the isolated enzyme in the reduced state. The Raman spectra of cytochrome oxidase isolated from bacterium, yeast, and bovine heart were different in the lower frequency region below 600 cm-1 but closely alike in the higher frequency region above 1200 cm-1.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D001419:Bacteria / Q000201:enzymology*; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D003576:Electron Transport Complex IV / Q000378:metabolism*; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology; D013045:Species Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods
D003576:Electron Transport Complex IV