Preparation, purification, and spectrophotometric characterization of cytochrome P450 1A2 conjugated with polyethylene glycol monomethyl ether.
Rabbit cytochrome P450 1A2 was modified with succinimidyl carbonate poly(ethylene glycol) monomethyl ether, purified by size exclusion high performance liquid chromatography, and lyophilized. Modification of cytochrome P450 1A2 caused no structural deformation of the heme as evidenced by the similarity of the spectral signatures for both the ferric form and the ferrous-CO complex to the respective forms for the unmodified enzyme. Ethoxyresorufin O-deethylation activity in the presence of iodosobenzene for the modified enzyme was comparable to that of the native enzyme.
Nakhgevany
R
R
Department of Chemistry, Northeastern University, Boston, Massachusetts 02115, USA.
Bonsu
O
O
Barber
J
J
Mabrouk
P A
PA
Ueng
Y F
YF
Bell
L C
LC
Guengerich
F P
FP
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D019388:Cytochrome P-450 CYP1A2; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005612:Freeze Drying; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology*; D010088:Oxidoreductases / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011092:Polyethylene Glycols; D011817:Rabbits; D013053:Spectrophotometry
C099846:succinimidyl carbonate poly(ethylene glycol) monomethyl ether; D011092:Polyethylene Glycols; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; D019388:Cytochrome P-450 CYP1A2