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Active site models of human cytochrome P450s based on NMR relaxation measurements

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Active site models of human cytochrome P450s based on NMR relaxation measurements. / Modi, S. ; Paine, M. J.; Oliver, C. F. ; Smith, G. ; Sutcliffe, M. J. ; Gilham, D. E. ; Shaw, A. N. J. ; Lian, L. Y. ; Primrose, W. U. ; Wolf, C. R. ; Roberts, G. C. K. .

In: FASEB Journal, Vol. 11, No. 9, 1997, p. A771-A771.

Research output: Contribution to journalMeeting abstract

Harvard

Modi, S, Paine, MJ, Oliver, CF, Smith, G, Sutcliffe, MJ, Gilham, DE, Shaw, ANJ, Lian, LY, Primrose, WU, Wolf, CR & Roberts, GCK 1997, 'Active site models of human cytochrome P450s based on NMR relaxation measurements' FASEB Journal, vol 11, no. 9, pp. A771-A771.

APA

Modi, S., Paine, M. J., Oliver, C. F., Smith, G., Sutcliffe, M. J., Gilham, D. E., Shaw, A. N. J., Lian, L. Y., Primrose, W. U., Wolf, C. R., & Roberts, G. C. K. (1997). Active site models of human cytochrome P450s based on NMR relaxation measurements. FASEB Journal, 11(9), A771-A771

Vancouver

Modi S, Paine MJ, Oliver CF, Smith G, Sutcliffe MJ, Gilham DE et al. Active site models of human cytochrome P450s based on NMR relaxation measurements. FASEB Journal. 1997;11(9):A771-A771.

Author

Modi, S. ; Paine, M. J.; Oliver, C. F. ; Smith, G. ; Sutcliffe, M. J. ; Gilham, D. E. ; Shaw, A. N. J. ; Lian, L. Y. ; Primrose, W. U. ; Wolf, C. R. ; Roberts, G. C. K. / Active site models of human cytochrome P450s based on NMR relaxation measurements.

In: FASEB Journal, Vol. 11, No. 9, 1997, p. A771-A771.

Research output: Contribution to journalMeeting abstract

Bibtex - Download

@article{b7b77eef899e470393b01d0625109ff2,
title = "Active site models of human cytochrome P450s based on NMR relaxation measurements",
author = "S. Modi and Paine, {M. J.} and Oliver, {C. F.} and G. Smith and Sutcliffe, {M. J.} and Gilham, {D. E.} and Shaw, {A. N. J.} and Lian, {L. Y.} and Primrose, {W. U.} and Wolf, {C. R.} and Roberts, {G. C. K.}",
year = "1997",
volume = "11",
number = "9",
pages = "A771--A771",
journal = "FASEB Journal",
issn = "0892-6638",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Active site models of human cytochrome P450s based on NMR relaxation measurements

A1 - Modi,S.

A1 - Paine,M. J.

A1 - Oliver,C. F.

A1 - Smith,G.

A1 - Sutcliffe,M. J.

A1 - Gilham,D. E.

A1 - Shaw,A. N. J.

A1 - Lian,L. Y.

A1 - Primrose,W. U.

A1 - Wolf,C. R.

A1 - Roberts,G. C. K.

AU - Modi,S.

AU - Paine,M. J.

AU - Oliver,C. F.

AU - Smith,G.

AU - Sutcliffe,M. J.

AU - Gilham,D. E.

AU - Shaw,A. N. J.

AU - Lian,L. Y.

AU - Primrose,W. U.

AU - Wolf,C. R.

AU - Roberts,G. C. K.

PY - 1997

Y1 - 1997

N2 - The various mammalian cytochrome P450s (2D6, 3A4. etc.) have been produced in large quantities by expression of their cDN As in baculovirus or e.coli. A poly-histidine extension has been incorporated at the C-terminus of the expressed proteins, which, after purification of the protein on a nickelagarose column, could be removed proteolytically by treatment with thrombin. The quantities produced allowed direct study of the interaction of substrates with P450s using NMR. We have studied the binding of the different substrates to the Cytochrome P450 2D6 and 3 A4 by measurements of the relaxation effects of the unpaired electron of the haem iron on the protons of the bound substrates'. Using these measurements on the substrate protons and the known X-ray crystal structures of four P450s, a model for the initial enzyme-substrate complex was built. Based on these results, mutants were designed to explore the substrate specificity of the enzyme. In Cytochrome P450 2D6, the F483I mutant was shown to accommodate larger substrate (like testosterone) as compared to wild type2.

AB - The various mammalian cytochrome P450s (2D6, 3A4. etc.) have been produced in large quantities by expression of their cDN As in baculovirus or e.coli. A poly-histidine extension has been incorporated at the C-terminus of the expressed proteins, which, after purification of the protein on a nickelagarose column, could be removed proteolytically by treatment with thrombin. The quantities produced allowed direct study of the interaction of substrates with P450s using NMR. We have studied the binding of the different substrates to the Cytochrome P450 2D6 and 3 A4 by measurements of the relaxation effects of the unpaired electron of the haem iron on the protons of the bound substrates'. Using these measurements on the substrate protons and the known X-ray crystal structures of four P450s, a model for the initial enzyme-substrate complex was built. Based on these results, mutants were designed to explore the substrate specificity of the enzyme. In Cytochrome P450 2D6, the F483I mutant was shown to accommodate larger substrate (like testosterone) as compared to wild type2.

UR - http://www.fasebj.org/

M1 - Meeting abstract

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 9

VL - 11

SP - A771-A771

ER -

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