Theoretical study of the product specificity in the hydroxylation of camphor, norcamphor, 5,5-difluorocamphor, and pericyclocamphanone by cytochrome P-450cam.
The hydroxylations of d-camphor, norcamphor, pericyclocamphanone, and 5,5-difluorocamphor by cytochrome P-450cam have been examined using theoretical methods to identify and characterize properties which determine product specificity. Experimental results indicate that each molecule is hydroxylated with quite different regio-specificity when metabolized by P-450cam. This result is surprising in view of their overall structural similiarity. Herein we report the results of calculations on d-camphor and three of its analogues which suggest that all of these molecules should, when metabolized by P-450cam, form hydroxylation products and predict the product distribution for each. Our conclusions are based on two fundamental criteria which are consistent with a generally accepted radical mechanism in determining product specificity in these molecules: 1) relative heats of formation of the radicals formed by abstracting a hydrogen, and 2) orientation of the substrate molecule with respect to the putative active oxygen species bound to iron. Our results explain the experimental observations for camphor and 5,5-difluorocamphor but disagree with original published results for norcamphor and pericyclocamphanone. In light of our results, new experiments have been performed for norcamphor and the original data reexamined for pericyclocamphanone. Our predictions have recently been experimentally confirmed for norcamphor, and unpublished data (Dr. S. Sligar) suggest that the same is true for pericyclocamphanone.
Collins
J R
JR
SRI International, Menlo Park, California 94025.
Loew
G H
GH
D001665:Binding Sites; D002164:Camphor / Q000031:analogs & derivatives* / Q000378:metabolism*; D019475:Camphor 5-Monooxygenase; D055598:Chemical Phenomena; D002621:Chemistry; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006860:Hydrogen Bonding; D006900:Hydroxylation; D007501:Iron / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009636:Norbornanes / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism; D013816:Thermodynamics
D009636:Norbornanes; C055059:norcamphor; D002164:Camphor; C020453:beta-pericyclocamphanone; D003577:Cytochrome P-450 Enzyme System; C043709:5,5-difluorocamphor; D007501:Iron; D006899:Mixed Function Oxygenases; D019475:Camphor 5-Monooxygenase; D010100:Oxygen