A mechanistic rationalisation for the substrate specificity of recombinant mammalian 4-hydroxyphenylpyruvate dioxygenase (4-HPPD)

Nicholas P. Crouch; Robert M. Adlington; Jack E. Baldwin; Meng Huee Lee; Colin H. MacKinnon

doi:10.1016/S0040-4020(97)00398-0

A mechanistic rationalisation for the substrate specificity of recombinant mammalian 4-hydroxyphenylpyruvate dioxygenase (4-HPPD)

Nicholas P. Crouch^*, Robert M. Adlington, Jack E. Baldwin, Meng Huee Lee, Colin H. MacKinnon

^*Corresponding author for this work

University of Oxford

Research output: Contribution to journal › Article › peer-review

50 Citations (Scopus)

Abstract

The isolation and purification of α-ketoisocaproate dioxygenase [α-KICD] from rat liver is described. Sequence determination of the purified protein revealed it to have complete homology to rat liver 4-hydroxyphenylpyruvate dioxygenase [4-HPPD] which was confirmed by the cloning and expression of the gene encoding 4-HPPD in E. coli. Examination of the substrate specificity of the resulting soluble recombinant protein revealed it to be capable of the oxidative decarboxylation of a range of ketoacids derived from proteinogenic amino acids. The significance of the turnover of these different ketoacids is discussed in relation to the mechanism of this fascinating enzyme.

Original language	English
Pages (from-to)	6993-7010
Number of pages	18
Journal	Tetrahedron
Volume	53
Issue number	20
DOIs	https://doi.org/10.1016/S0040-4020(97)00398-0
Publication status	Published - 19 May 1997
Externally published	Yes

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@article{0aa7624344934624be191470a9ffa4be,

title = "A mechanistic rationalisation for the substrate specificity of recombinant mammalian 4-hydroxyphenylpyruvate dioxygenase (4-HPPD)",

abstract = "The isolation and purification of α-ketoisocaproate dioxygenase [α-KICD] from rat liver is described. Sequence determination of the purified protein revealed it to have complete homology to rat liver 4-hydroxyphenylpyruvate dioxygenase [4-HPPD] which was confirmed by the cloning and expression of the gene encoding 4-HPPD in E. coli. Examination of the substrate specificity of the resulting soluble recombinant protein revealed it to be capable of the oxidative decarboxylation of a range of ketoacids derived from proteinogenic amino acids. The significance of the turnover of these different ketoacids is discussed in relation to the mechanism of this fascinating enzyme.",

author = "Crouch, {Nicholas P.} and Adlington, {Robert M.} and Baldwin, {Jack E.} and Lee, {Meng Huee} and MacKinnon, {Colin H.}",

note = "Funding Information: We thank the EPSRC for a grant (to CHM) and the British Council for an Overseas Research studentship (to MHL). We thank also Prof. R. A. Pascal, Princeton University, USA, for help in the early stages of our work, Dr Y. Fujishima and Mr J.P.N. Pitt for assistance with protein purification, Dr A. C. Willis for peptide sequencing, Dr. G. J. Pritchard for help with the synthetic chemistry and Dr Z. H. Zhang for assistance in the cloning of mammalian 4-HPPD.",

year = "1997",

month = may,

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doi = "10.1016/S0040-4020(97)00398-0",

language = "English",

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pages = "6993--7010",

journal = "Tetrahedron",

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TY - JOUR

T1 - A mechanistic rationalisation for the substrate specificity of recombinant mammalian 4-hydroxyphenylpyruvate dioxygenase (4-HPPD)

AU - Crouch, Nicholas P.

AU - Adlington, Robert M.

AU - Baldwin, Jack E.

AU - Lee, Meng Huee

AU - MacKinnon, Colin H.

N1 - Funding Information: We thank the EPSRC for a grant (to CHM) and the British Council for an Overseas Research studentship (to MHL). We thank also Prof. R. A. Pascal, Princeton University, USA, for help in the early stages of our work, Dr Y. Fujishima and Mr J.P.N. Pitt for assistance with protein purification, Dr A. C. Willis for peptide sequencing, Dr. G. J. Pritchard for help with the synthetic chemistry and Dr Z. H. Zhang for assistance in the cloning of mammalian 4-HPPD.

PY - 1997/5/19

Y1 - 1997/5/19

N2 - The isolation and purification of α-ketoisocaproate dioxygenase [α-KICD] from rat liver is described. Sequence determination of the purified protein revealed it to have complete homology to rat liver 4-hydroxyphenylpyruvate dioxygenase [4-HPPD] which was confirmed by the cloning and expression of the gene encoding 4-HPPD in E. coli. Examination of the substrate specificity of the resulting soluble recombinant protein revealed it to be capable of the oxidative decarboxylation of a range of ketoacids derived from proteinogenic amino acids. The significance of the turnover of these different ketoacids is discussed in relation to the mechanism of this fascinating enzyme.

AB - The isolation and purification of α-ketoisocaproate dioxygenase [α-KICD] from rat liver is described. Sequence determination of the purified protein revealed it to have complete homology to rat liver 4-hydroxyphenylpyruvate dioxygenase [4-HPPD] which was confirmed by the cloning and expression of the gene encoding 4-HPPD in E. coli. Examination of the substrate specificity of the resulting soluble recombinant protein revealed it to be capable of the oxidative decarboxylation of a range of ketoacids derived from proteinogenic amino acids. The significance of the turnover of these different ketoacids is discussed in relation to the mechanism of this fascinating enzyme.

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U2 - 10.1016/S0040-4020(97)00398-0

DO - 10.1016/S0040-4020(97)00398-0

M3 - Article

AN - SCOPUS:0030952515

SN - 0040-4020

VL - 53

SP - 6993

EP - 7010

JO - Tetrahedron

JF - Tetrahedron

IS - 20

ER -

A mechanistic rationalisation for the substrate specificity of recombinant mammalian 4-hydroxyphenylpyruvate dioxygenase (4-HPPD)

Abstract

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