Recombinant cytochrome c peroxidase folds properly without conformational "annealing".
Recombinant cytochrome c peroxidase isolated from Escherichia coli has recently been reported to exhibit an abnormal electronic absorption spectrum that is converted to the normal spectrum after conformational "annealing" of the recombinant enzyme by passage over a cytochrome c affinity column. The current report provides evidence that the abnormal spectrum observed in some preparations of recombinant cytochrome c peroxidase arises from the presence of contaminant, damaged forms cytochrome c peroxidase with altered spectra. Removal of these contaminant forms produces a major cytochrome c peroxidase fraction with a normal spectrum. We conclude that elution of recombinant cytochrome c peroxidase over a cytochrome c affinity column does not produce normal enzyme through conformational "annealing" but that it produces purified enzyme through removal of contaminants.
Ferrer
J C
JC
Department of Biochemistry, University of British Columbia, Vancouver, Canada.
Ring
M
M
Mauk
A G
AG
D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013053:Spectrophotometry
D011994:Recombinant Proteins; D003576:Electron Transport Complex IV