Capturing enzyme structure prior to reaction initiation: Tropinone reductase-II-substrate complexes

Atsuko Yamashita, Masaharu Endo, Tsuneyuki Higashi, Toru Nakatsu, Yasuyuki Yamada, Jun'ichi Oda, Hiroaki Kato

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

To understand the catalytic mechanism of an enzyme, it is crucial to determine the crystallographic structures corresponding to the individual reaction steps. Here, we report two crystal structures of enzyme - substrate complexes prior to reaction initiation: tropinone reductase-II (TR-II) NADPH and TR-II - NADPH - tropinone complexes, determined from the identical crystals. A combination of two kinetic crystallographic techniques, a continuous flow of the substrates and Laue diffraction measurements, enabled us to capture the transit structures prior to the reaction proceeding. A structure comparison of the enzyme - substrate complex elucidated in this study with the enzyme-product complex in our previous study indicates that one of the substrates, tropinone, is rotated relative to the product so as to make the spatial organization in the active site favorable for the reaction to proceed. Side chains of the residues in the active site also alter their conformations to keep the complementarity of the space for the substrate or the product and to assist the rotational movement.

Original languageEnglish
Pages (from-to)5566-5573
Number of pages8
JournalBiochemistry
Volume42
Issue number19
DOIs
Publication statusPublished - May 20 2003
Externally publishedYes

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tropinone reductase
Substrates
Enzymes
NADP
Catalytic Domain
Conformations
Diffraction
Crystal structure
Crystals

ASJC Scopus subject areas

  • Biochemistry

Cite this

Capturing enzyme structure prior to reaction initiation : Tropinone reductase-II-substrate complexes. / Yamashita, Atsuko; Endo, Masaharu; Higashi, Tsuneyuki; Nakatsu, Toru; Yamada, Yasuyuki; Oda, Jun'ichi; Kato, Hiroaki.

In: Biochemistry, Vol. 42, No. 19, 20.05.2003, p. 5566-5573.

Research output: Contribution to journalArticle

Yamashita, A, Endo, M, Higashi, T, Nakatsu, T, Yamada, Y, Oda, J & Kato, H 2003, 'Capturing enzyme structure prior to reaction initiation: Tropinone reductase-II-substrate complexes', Biochemistry, vol. 42, no. 19, pp. 5566-5573. https://doi.org/10.1021/bi0272712
Yamashita, Atsuko ; Endo, Masaharu ; Higashi, Tsuneyuki ; Nakatsu, Toru ; Yamada, Yasuyuki ; Oda, Jun'ichi ; Kato, Hiroaki. / Capturing enzyme structure prior to reaction initiation : Tropinone reductase-II-substrate complexes. In: Biochemistry. 2003 ; Vol. 42, No. 19. pp. 5566-5573.
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