(+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase

A T Dinkova-Kostova; D R Gang; L B Davin; D L Bedgar; A Chu; N G Lewis

doi:10.1074/jbc.271.46.29473

Back

(+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase

Journal article

Open access

Peer reviewed

(+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase

A T Dinkova-Kostova, D R Gang, L B Davin, D L Bedgar, A Chu and N G Lewis

The Journal of biological chemistry, Vol.271(46), pp.29473-29482

11/15/1996

DOI: https://doi.org/10.1074/jbc.271.46.29473

Handle:

https://hdl.handle.net/2376/114251

PMID: 8910615

Files and links (2)

pdf

1-s2.0-S0021925818350907-main436.27 kBDownload View

CC BY V4.0, Open Access

url

https://doi.org/10.1074/jbc.271.46.29473View

Published (Version of record) Open

Abstract

Amino Acid Sequence

Recombinant Fusion Proteins - isolation & purification

Chromatography, Ion Exchange

Temperature

Molecular Weight

Oxidoreductases - metabolism

Oxidoreductases - genetics

Electrophoresis, Polyacrylamide Gel

Molecular Sequence Data

Recombinant Fusion Proteins - metabolism

Plants - enzymology

Sequence Homology, Amino Acid

Oxidoreductases - isolation & purification

Chromatography, Affinity

Escherichia coli - genetics

Base Sequence

Isoelectric Point

Cloning, Molecular

Recombinant Fusion Proteins - genetics

Oxidoreductases Acting on CH-CH Group Donors

Kinetics

DNA, Complementary

Hydrogen-Ion Concentration

Lignans are a widely distributed class of natural products, whose functions and distribution suggest that they are one of the earliest forms of defense to have evolved in vascular plants; some, such as podophyllotoxin and enterodiol, have important roles in cancer chemotherapy and prevention, respectively. Entry into lignan enzymology has been gained by the approximately 3000-fold purification of two isoforms of (+)-pinoresinol/(+)-lariciresinol reductase, a pivotal branchpoint enzyme in lignan biosynthesis. Both have comparable ( approximately 34.9 kDa) molecular mass and kinetic (Vmax/Km) properties and catalyze sequential, NADPH-dependent, stereospecific, hydride transfers where the incoming hydride takes up the pro-R position. The gene encoding (+)-pinoresinol/(+)-lariciresinol reductase has been cloned and the recombinant protein heterologously expressed as a functional beta-galactosidase fusion protein. Its amino acid sequence reveals a strong homology to isoflavone reductase, a key branchpoint enzyme in isoflavonoid metabolism and primarily found in the Fabaceae (angiosperms). This is of great evolutionary significance since both lignans and isoflavonoids have comparable plant defense properties, as well as similar roles as phytoestrogens. Given that lignans are widespread from primitive plants onwards, whereas the isoflavone reductase-derived isoflavonoids are mainly restricted to the Fabaceae, it is tempting to speculate that this branch of the isoflavonoid pathway arose via evolutionary divergence from that giving the lignans.

Metrics

10 Record Views

See more details

Details

Title: (+)-Pinoresinol/(+)-lariciresinol reductase from Forsythia intermedia. Protein purification, cDNA cloning, heterologous expression and comparison to isoflavone reductase
Creators: A T Dinkova-Kostova - Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA. lewisn@wsu.edu
D R Gang
L B Davin
D L Bedgar
A Chu
N G Lewis
Publication Details: The Journal of biological chemistry, Vol.271(46), pp.29473-29482
Academic Unit: Biological Chemistry, Institute of
Publisher: United States
Identifiers: 99900547715901842
Language: English
Resource Type: Journal article