Spectroelectrochemistry of cytochrome P450cam.
The spectroelectrochemistry of camphor-bound cytochrome P450cam (P450cam) using gold electrodes is described. The electrodes were modified with either 4,4(')-dithiodipyridin or sodium dithionite. Electrolysis of P450cam was carried out when the enzyme was in solution, while at the same time UV-visible absorption spectra were recorded. Reversible oxidation and reduction could be observed with both 4,4(')-dithiodipyridin and dithionite modified electrodes. A formal potential (E(0')) of -373mV vs Ag/AgCl 1M KCl was determined. The spectra of P450cam complexed with either carbon monoxide or metyrapone, both being inhibitors of P450 catalysis, clearly indicated that the protein retained its native state in the electrochemical cell during electrolysis.
Bistolas
Nikitas
N
Department of Analytical Biochemistry, University of Potsdam, Karl-Liebknecht-Str. 24-25, 14476, Golm, Germany.
Christenson
Andreas
A
Ruzgas
Tautgirdas
T
Jung
Christiane
C
Scheller
Frieder W
FW
Wollenberger
Ulla
U
D002164:Camphor / Q000737:chemistry / Q000378:metabolism; D019475:Camphor 5-Monooxygenase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D002248:Carbon Monoxide / Q000737:chemistry / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry; D004227:Dithionite / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D008797:Metyrapone / Q000494:pharmacology; D010084:Oxidation-Reduction; D016958:Pseudomonas putida / Q000201:enzymology; D011725:Pyridines / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000295:instrumentation / Q000379:methods*
D004220:Disulfides; D004791:Enzyme Inhibitors; D011725:Pyridines; D004227:Dithionite; C007542:4,4'-dipyridyl disulfide; D006046:Gold; D002164:Camphor; D002248:Carbon Monoxide; D019475:Camphor 5-Monooxygenase; D008797:Metyrapone