Structures of recombinant human and mouse NAD(P)H: quinone oxidoreductases: Species comparison and structural changes with substrate binding and release

Margarita Faig, Mario Antonio Bianchet, Paul Talalay, Shiuan Chen, Shannon Winski, David Ross, Mario L Amzel

Research output: Contribution to journalArticle

Abstract

NAD(P)H/quinone acceptor oxidoreductase (QR1, NQO1, formerly DT- diaphorase; EC 1.6.99.2) protects animal cells from the deleterious and carcinogenic effects of quinones and other electrophiles. In this paper we report the apoenzyme structures of human (at 1.7-Å resolution) and mouse (2.8 Å) QR1 and the complex of the human enzyme with the substrate duroquinone (2.5 Å) (2,3,5,6-tetramethyl-p-benzoquinone). In addition to providing a description and rationale of the structural and catalytic differences among several species, these structures reveal the changes that accompany substrate or cofactor (NAD) binding and release. Tyrosine-128 and the loop spanning residues 232-236 close the binding site, partially occupying the space left vacant by the departing molecule (substrate or cofactor). These changes highlight the exquisite control of access to the catalytic site that is required by the ping-pong mechanism in which, after reducing the flavin, NAD(P)+ leaves the catalytic site and allows substrate to bind at the vacated position. In the human QR1-duroquinone structure one ring carbon is significantly closer to the flavin NS, suggesting a direct hydride transfer to this atom.

Original languageEnglish (US)
Pages (from-to)3177-3182
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume97
Issue number7
DOIs
StatePublished - Mar 28 2000

Fingerprint

NAD
NAD(P)H Dehydrogenase (Quinone)
Oxidoreductases
Catalytic Domain
Apoenzymes
Quinones
Tyrosine
Carbon
Binding Sites
Enzymes
benzoquinone
4,6-dinitro-o-cresol
duroquinone

Keywords

  • Cancer
  • Chemoprotection
  • Chemotherapy
  • DT diaphorase

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

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title = "Structures of recombinant human and mouse NAD(P)H: quinone oxidoreductases: Species comparison and structural changes with substrate binding and release",
abstract = "NAD(P)H/quinone acceptor oxidoreductase (QR1, NQO1, formerly DT- diaphorase; EC 1.6.99.2) protects animal cells from the deleterious and carcinogenic effects of quinones and other electrophiles. In this paper we report the apoenzyme structures of human (at 1.7-{\AA} resolution) and mouse (2.8 {\AA}) QR1 and the complex of the human enzyme with the substrate duroquinone (2.5 {\AA}) (2,3,5,6-tetramethyl-p-benzoquinone). In addition to providing a description and rationale of the structural and catalytic differences among several species, these structures reveal the changes that accompany substrate or cofactor (NAD) binding and release. Tyrosine-128 and the loop spanning residues 232-236 close the binding site, partially occupying the space left vacant by the departing molecule (substrate or cofactor). These changes highlight the exquisite control of access to the catalytic site that is required by the ping-pong mechanism in which, after reducing the flavin, NAD(P)+ leaves the catalytic site and allows substrate to bind at the vacated position. In the human QR1-duroquinone structure one ring carbon is significantly closer to the flavin NS, suggesting a direct hydride transfer to this atom.",
keywords = "Cancer, Chemoprotection, Chemotherapy, DT diaphorase",
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T2 - quinone oxidoreductases: Species comparison and structural changes with substrate binding and release

AU - Faig, Margarita

AU - Bianchet, Mario Antonio

AU - Talalay, Paul

AU - Chen, Shiuan

AU - Winski, Shannon

AU - Ross, David

AU - Amzel, Mario L

PY - 2000/3/28

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N2 - NAD(P)H/quinone acceptor oxidoreductase (QR1, NQO1, formerly DT- diaphorase; EC 1.6.99.2) protects animal cells from the deleterious and carcinogenic effects of quinones and other electrophiles. In this paper we report the apoenzyme structures of human (at 1.7-Å resolution) and mouse (2.8 Å) QR1 and the complex of the human enzyme with the substrate duroquinone (2.5 Å) (2,3,5,6-tetramethyl-p-benzoquinone). In addition to providing a description and rationale of the structural and catalytic differences among several species, these structures reveal the changes that accompany substrate or cofactor (NAD) binding and release. Tyrosine-128 and the loop spanning residues 232-236 close the binding site, partially occupying the space left vacant by the departing molecule (substrate or cofactor). These changes highlight the exquisite control of access to the catalytic site that is required by the ping-pong mechanism in which, after reducing the flavin, NAD(P)+ leaves the catalytic site and allows substrate to bind at the vacated position. In the human QR1-duroquinone structure one ring carbon is significantly closer to the flavin NS, suggesting a direct hydride transfer to this atom.

AB - NAD(P)H/quinone acceptor oxidoreductase (QR1, NQO1, formerly DT- diaphorase; EC 1.6.99.2) protects animal cells from the deleterious and carcinogenic effects of quinones and other electrophiles. In this paper we report the apoenzyme structures of human (at 1.7-Å resolution) and mouse (2.8 Å) QR1 and the complex of the human enzyme with the substrate duroquinone (2.5 Å) (2,3,5,6-tetramethyl-p-benzoquinone). In addition to providing a description and rationale of the structural and catalytic differences among several species, these structures reveal the changes that accompany substrate or cofactor (NAD) binding and release. Tyrosine-128 and the loop spanning residues 232-236 close the binding site, partially occupying the space left vacant by the departing molecule (substrate or cofactor). These changes highlight the exquisite control of access to the catalytic site that is required by the ping-pong mechanism in which, after reducing the flavin, NAD(P)+ leaves the catalytic site and allows substrate to bind at the vacated position. In the human QR1-duroquinone structure one ring carbon is significantly closer to the flavin NS, suggesting a direct hydride transfer to this atom.

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