Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

Tongpil Min; Hiroyuki Kasahara; Diana L Bedgar; Buhyun Youn; Paulraj K Lawrence; David R Gang; Steven C Halls; HaJeung Park; Jacqueline L Hilsenbeck; Laurence B Davin; Norman G Lewis; ChulHee Kang

doi:10.1074/jbc.M308493200

Back

Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

Journal article

Open access

Peer reviewed

Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

Tongpil Min, Hiroyuki Kasahara, Diana L Bedgar, Buhyun Youn, Paulraj K Lawrence, David R Gang, Steven C Halls, HaJeung Park, Jacqueline L Hilsenbeck, Laurence B Davin, …

The Journal of biological chemistry, Vol.278(50), pp.50714-50723

12/12/2003

DOI: https://doi.org/10.1074/jbc.M308493200

Handle:

https://hdl.handle.net/2376/115493

PMID: 13129921

Files and links (1)

url

https://doi.org/10.1074/jbc.M308493200View

Published (Version of record) Open

Abstract

Protein Structure, Tertiary

Amino Acid Sequence

Mutagenesis, Site-Directed

Models, Chemical

Protein Structure, Secondary

Lignans - chemistry

Models, Molecular

Molecular Sequence Data

Crystallography, X-Ray

Ultracentrifugation

NADP - chemistry

Oxidoreductases - chemistry

Plasmids - metabolism

Oxidoreductases Acting on CH-CH Group Donors - chemistry

Sequence Homology, Amino Acid

Escherichia coli - metabolism

Protein Binding

Protein Conformation

Furans

Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

Metrics

8 Record Views

See more details

Details

Title: Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases
Creators: Tongpil Min - School of Molecular Biosciences, Washington State University, Pullman, Washington 99164-4660, USA
Hiroyuki Kasahara
Diana L Bedgar
Buhyun Youn
Paulraj K Lawrence
David R Gang
Steven C Halls
HaJeung Park
Jacqueline L Hilsenbeck
Laurence B Davin
Norman G Lewis
ChulHee Kang
Publication Details: The Journal of biological chemistry, Vol.278(50), pp.50714-50723
Academic Unit: Chemistry, Department of; Biological Chemistry, Institute of
Publisher: United States
Identifiers: 99900547685201842
Language: English
Resource Type: Journal article