Structure of tropinone reductase-II complexed with NADP+ and pseudotropine at 1.9 Å resolution

Implication for stereospecific substrate binding and catalysis

Atsuko Yamashita, Hiroaki Kato, Soichi Wakatsuki, Takashi Tomizaki, Toru Nakatsu, Keiji Nakajima, Takashi Hashimoto, Yasuyuki Yamada, Jun'ichi Oda

Research output: Contribution to journalArticle

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Abstract

Tropinone reductase-II (TR-II) catalyzes the NADPH-dependent reduction of the carbonyl group of tropinone to a β-hydroxyl group. The crystal structure of TR-II complexed with NADP+ and pseudotropine (Ψ-tropine) has been determined at 1.9 Å resolution. A seven-residue peptide near the active site, disordered in the unliganded structure, is fixed in the ternary complex by participation of the cofactor and substrate binding. The Ψ-tropine molecule is bound in an orientation which satisfies the product configuration and the stereochemical arrangement toward the cofactor. The substrate binding site displays a complementarity to the bound substrate (Ψ-tropine) in its correct orientation. In addition, electrostatic interactions between the substrate and Glu156 seem to specify the binding position and orientation of the substrate. A comparison between the active sites in TR-II and TR-I shows that they provide different van der Waals surfaces and electrostatic features. These differences likely contribute to the correct binding mode of the substrates, which are in opposite orientations in TR-II and TR-I, and to different reaction stereospecificities. The active site structure in the TR- II ternary complex also suggests that the arrangement of the substrate, cofactor, and catalytic residues is stereoelectronically favorable for the reaction.

Original languageEnglish
Pages (from-to)7630-7637
Number of pages8
JournalBiochemistry
Volume38
Issue number24
DOIs
Publication statusPublished - Jun 15 1999
Externally publishedYes

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tropinone reductase
Catalysis
NADP
Catalytic Domain
Substrates
Static Electricity
Hydroxyl Radical
Binding Sites
Peptides
Coulomb interactions
tropine
Electrostatics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structure of tropinone reductase-II complexed with NADP+ and pseudotropine at 1.9 Å resolution : Implication for stereospecific substrate binding and catalysis. / Yamashita, Atsuko; Kato, Hiroaki; Wakatsuki, Soichi; Tomizaki, Takashi; Nakatsu, Toru; Nakajima, Keiji; Hashimoto, Takashi; Yamada, Yasuyuki; Oda, Jun'ichi.

In: Biochemistry, Vol. 38, No. 24, 15.06.1999, p. 7630-7637.

Research output: Contribution to journalArticle

Yamashita, Atsuko ; Kato, Hiroaki ; Wakatsuki, Soichi ; Tomizaki, Takashi ; Nakatsu, Toru ; Nakajima, Keiji ; Hashimoto, Takashi ; Yamada, Yasuyuki ; Oda, Jun'ichi. / Structure of tropinone reductase-II complexed with NADP+ and pseudotropine at 1.9 Å resolution : Implication for stereospecific substrate binding and catalysis. In: Biochemistry. 1999 ; Vol. 38, No. 24. pp. 7630-7637.
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abstract = "Tropinone reductase-II (TR-II) catalyzes the NADPH-dependent reduction of the carbonyl group of tropinone to a β-hydroxyl group. The crystal structure of TR-II complexed with NADP+ and pseudotropine (Ψ-tropine) has been determined at 1.9 {\AA} resolution. A seven-residue peptide near the active site, disordered in the unliganded structure, is fixed in the ternary complex by participation of the cofactor and substrate binding. The Ψ-tropine molecule is bound in an orientation which satisfies the product configuration and the stereochemical arrangement toward the cofactor. The substrate binding site displays a complementarity to the bound substrate (Ψ-tropine) in its correct orientation. In addition, electrostatic interactions between the substrate and Glu156 seem to specify the binding position and orientation of the substrate. A comparison between the active sites in TR-II and TR-I shows that they provide different van der Waals surfaces and electrostatic features. These differences likely contribute to the correct binding mode of the substrates, which are in opposite orientations in TR-II and TR-I, and to different reaction stereospecificities. The active site structure in the TR- II ternary complex also suggests that the arrangement of the substrate, cofactor, and catalytic residues is stereoelectronically favorable for the reaction.",
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