Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101)

Michael Strickler; Barry M Goldstein; Kathleen Maxfield; Laura Shireman; Gyungyoun Kim; Donald S Matteson; Jeffrey P Jones

doi:10.1021/bi034833o

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Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101)

Journal article

Peer reviewed

Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101)

Michael Strickler, Barry M Goldstein, Kathleen Maxfield, Laura Shireman, Gyungyoun Kim, Donald S Matteson and Jeffrey P Jones

Biochemistry (Easton), Vol.42(41), pp.11943-11950

10/21/2003

DOI: https://doi.org/10.1021/bi034833o

Handle:

https://hdl.handle.net/2376/114084

PMID: 14556625

Abstract

Recombinant Proteins - metabolism

Models, Chemical

Crystallization

Models, Molecular

Recombinant Proteins - chemistry

Substrate Specificity

Crystallography, X-Ray

Carbon Monoxide - chemistry

Carbon Monoxide - metabolism

Recombinant Proteins - isolation & purification

Nicotine - metabolism

Camphor 5-Monooxygenase - chemistry

Computer Simulation

Nicotine - chemistry

Camphor 5-Monooxygenase - metabolism

Camphor 5-Monooxygenase - isolation & purification

Spectrophotometry

Binding Sites

Crystallographic and spectroscopic studies have been undertaken to characterize the binding behavior of the non-native substrate nicotine in the active site of the monooxygenase hemoprotein cytochrome P450cam. Despite the existence of a theoretical model that is consistent with the observed distribution of monooxygenation products, the crystal structure of the complex indicates that the primary binding mode of nicotine is unproductive. The structure is confirmed by spectral data that indicate direct coordination of substrate pyridine nitrogen with the heme iron. This would be the proper structure for evaluating binding affinity and inhibition. Reduction of the heme from Fe(III) to Fe(II) and introduction of carbon monoxide into crystals of the nicotine-P450cam complex, to simulate molecular oxygen binding, produces reorientation of the nicotine. This orientation is the appropriate one for predicting regioselectivity and the kinetic features of substrate oxidation. While it is not clear that such complicated behavior will be exhibited for other enzyme-substrate interactions, it is clear that a single crystal structure for a given substrate-enzyme interaction may not provide a good description of the binding mode responsible for product formation.

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Title: Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101)
Creators: Michael Strickler - Department of Biophysics, University of Rochester, Rochester, New York 14627, USA
Barry M Goldstein
Kathleen Maxfield
Laura Shireman
Gyungyoun Kim
Donald S Matteson
Jeffrey P Jones
Publication Details: Biochemistry (Easton), Vol.42(41), pp.11943-11950
Academic Unit: Department of Chemistry
Publisher: United States
Grant note: RR-01646 / NCRR NIH HHS NIEHS 009122 / NCEH CDC HHS GM 32165 / NIGMS NIH HHS
Identifiers: 99900548388001842
Language: English
Resource Type: Journal article

Crystallographic studies on the complex behavior of nicotine binding to P450cam (CYP101)

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