A multisubstrate reductase from Plantago major: structure-function in the short chain reductase superfamily

Rachel Fellows, Christopher M. Russo, Catarina S. Silva, Soon Goo Lee, Joseph M. Jez, John D Chisholm, Chloe Zubieta, Max H. Nanao

Research output: Contribution to journalArticle

Abstract

The short chain dehydrogenase/reductase superfamily (SDR) is a large family of NAD(P)H-dependent enzymes found in all kingdoms of life. SDRs are particularly well-represented in plants, playing diverse roles in both primary and secondary metabolism. In addition, some plant SDRs are also able to catalyse a reductive cyclisation reaction critical for the biosynthesis of the iridoid backbone that contains a fused 5 and 6-membered ring scaffold. Mining the EST database of Plantago major, a medicinal plant that makes iridoids, we identified a putative 5β-progesterone reductase gene, PmMOR (P. major multisubstrate oxido-reductase), that is 60% identical to the iridoid synthase gene from Catharanthus roseus. The PmMOR protein was recombinantly expressed and its enzymatic activity assayed against three putative substrates, 8-oxogeranial, citral and progesterone. The enzyme demonstrated promiscuous enzymatic activity and was able to not only reduce progesterone and citral, but also to catalyse the reductive cyclisation of 8-oxogeranial. The crystal structures of PmMOR wild type and PmMOR mutants in complex with NADP+ or NAD+ and either 8-oxogeranial, citral or progesterone help to reveal the substrate specificity determinants and catalytic machinery of the protein. Site-directed mutagenesis studies were performed and provide a foundation for understanding the promiscuous activity of the enzyme.

Original languageEnglish (US)
Article number14796
JournalScientific Reports
Volume8
Issue number1
DOIs
StatePublished - Dec 1 2018

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citral
Plantago
Oxidoreductases
Progesterone
Cyclization
NAD
Progesterone Reductase
Enzymes
Iridoids
Catharanthus
Secondary Metabolism
Expressed Sequence Tags
Medicinal Plants
Substrate Specificity
Site-Directed Mutagenesis
NADP
Genes
Proteins
short chain trans-2-enoyl-CoA reductase
Databases

ASJC Scopus subject areas

  • General

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A multisubstrate reductase from Plantago major : structure-function in the short chain reductase superfamily. / Fellows, Rachel; Russo, Christopher M.; Silva, Catarina S.; Lee, Soon Goo; Jez, Joseph M.; Chisholm, John D; Zubieta, Chloe; Nanao, Max H.

In: Scientific Reports, Vol. 8, No. 1, 14796, 01.12.2018.

Research output: Contribution to journalArticle

Fellows, Rachel ; Russo, Christopher M. ; Silva, Catarina S. ; Lee, Soon Goo ; Jez, Joseph M. ; Chisholm, John D ; Zubieta, Chloe ; Nanao, Max H. / A multisubstrate reductase from Plantago major : structure-function in the short chain reductase superfamily. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
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