title: Progesterone 5β-reductase genes of the Brassicaceae family as function-associated molecular markers.
abstract: This study aimed to define progesterone 5β-reductases (P5βR, EC 1.3.99.6, enone 1,4-reductases) as function-associated molecular markers at the plant family level. Therefore cDNAs were isolated from 25 Brassicaceae species, including two species, Erysimum crepidifolium and Draba aizoides, known to produce cardiac glycosides. The sequences were used in a molecular phylogeny study. The cladogram created is congruent to the existing molecular analyses. Recombinant His-tagged forms of the P5βR cDNAs from Aethionema grandiflorum, Draba aizoides, Nasturtium officinale, Raphanus sativus and Sisymbrium officinale were expressed in E. coli. Enone 1,4-reductase activity was demonstrated in vitro using progesterone and 2-cyclohexen-1-one as substrates. Evidence is provided that functional P5βRs are ubiquitous in the Brassicaceae. The recombinant P5βR enzymes showed different substrate preferences towards progesterone and 2-cyclohexen-1-one. Sequence comparison of the catalytic pocket of the P5βR enzymes and homology modelling using Digitalis lanata P5βR (PDB ID: 2V6G) as template highlighted the importance of the hydrophobicity of the binding pocket for substrate discrimination. It is concluded that P5βR genes or P5βR proteins can be used as valuable function-associated molecular markers to infer taxonomic relationship and evolutionary diversification from a metabolic/catalytic perspective.
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Munkert
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: C
authors.initials: C
authors.lastname: Costa
authors.affiliation: University of the Academy of Sciences of Moldova, Chisinau, Moldova Republic.
authors.forename: O
authors.initials: O
authors.lastname: Budeanu
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Petersen
authors.affiliation: Agriculture Department, Universidade Federal de Lavras, Lavras, Brazil.
authors.forename: S
authors.initials: S
authors.lastname: Bertolucci
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Fischer
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: F
authors.initials: F
authors.lastname: Müller-Uri
authors.affiliation: Lehrstuhl für Pharmazeutische Biologie, Department Biologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Erlangen, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Kreis
mesh_terms: D000595:Amino Acid Sequence; D019607:Brassicaceae / Q000201:enzymology / Q000235:genetics*; D003512:Cyclohexanones / Q000378:metabolism; D018076:DNA, Complementary; D004070:Digitalis; D004926:Escherichia coli; D015870:Gene Expression; D017343:Genes, Plant; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D011375:Progesterone Reductase / Q000235:genetics* / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D003512:Cyclohexanones; D018076:DNA, Complementary; D010940:Plant Proteins; C028215:2-cyclohexen-1-one; D011374:Progesterone; D010088:Oxidoreductases; D011375:Progesterone Reductase; C090493:progesterone 5 beta-reductase
keywords: Brassicaceae; VEP1; enone 5β-reductase; metabolism; phylogeny