PDBsum entry 5mlr

Oxidoreductase

PDB id: 5mlr

Contents

Protein chain

364 a.a.

Ligands

GRQ ×2

NAP

Metals

_CL

_NA ×2

Waters ×525

PDB id:

5mlr

Name:

Oxidoreductase

Title:

Plantago major multifunctional oxidoreductase v150m mutant in complex with citral and NADP+

Structure:

Progesterone 5-beta-reductase. Chain: a. Engineered: yes

Source:

Plantago major. Common plantain. Organism_taxid: 29818. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008

Resolution:

1.46Å R-factor: 0.153 R-free: 0.169

Authors:

R.Fellows,C.M.Russo,S.G.Lee,J.M.Jez,J.D.Chisholm,C.Zubieta,M.Nanao

Key ref:

R.Fellows et al. (2018). A multisubstrate reductase from Plantago major: structure-function in the short chain reductase superfamily. Sci Rep, 8, 14796. PubMed id: 30287897

Date:

07-Dec-16 Release date: 01-Aug-18

Protein chain

D6N9X1  (D6N9X1_PLAMJ) -  Progesterone 5-beta-reductase from Plantago major

Seq: 389 a.a.

Struc: 364 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.1.1.1.145 - 3beta-hydroxy-Delta(5)-steroid dehydrogenase.
• Reaction: a 3beta-hydroxy-Delta₅-steroid + NAD⁺ = a 3-oxo-Delta₅-steroid + NADH + H⁺

3beta-hydroxy-Delta(5)-steroid (Bound ligand (Het Group name = GRQ) matches with 55.00% similarity) + NAD(+) (Bound ligand (Het Group name = NAP) matches with 91.67% similarity) = 3-oxo-Delta(5)-steroid + NADH + H(+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

Sci Rep 8:14796 (2018)

PubMed id: 30287897

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

R.Fellows, C.M.Russo, C.S.Silva, S.G.Lee, J.M.Jez, J.D.Chisholm, C.Zubieta, M.H.Nanao.

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.