title: Isosteric probes provide structural requirements essential for detoxification of the phytoalexin brassinin by the fungal pathogen Leptosphaeria maculans.
abstract: Brassinin is a plant defense metabolite with antimicrobial activity produced de novo by a variety of Brassica species in response to stress, that is, a phytoalexin. The inhibition of brassinin oxidase (BO), a brassinin-detoxifying enzyme produced by the phytopathogenic fungus Leptosphaeria maculans, is a target in our continuing search for novel crop protection agents. To probe the substrate specificity of BO, in particular the mechanism of the detoxification step, several analogues of brassinin, including functional group isosteres ((mono/dithio)carbamate, urea, and thiourea) and homologue methyl tryptaminedithiocarbamate, were investigated using fungal cultures and purified BO. It was concluded that the essential structural features of substrates of BO were: (i) an -NH at the (mono/dithio)carbamate, urea or thiourea group; (ii) a methylene bridge between indole and the functional group; (iii) a methyl or ethyl group attached to the thiol moiety of the (mono/di)thiocarbamate group. A general stepwise pathway for the oxidation of brassinin was proposed that accounts for the structural requirements of detoxification of brassinin analogues in L. maculans. All compounds that were BO substrates appeared to be oxidized in mycelial cultures to aldehydes, except for the two most polar compounds N'-(3-indolylmethyl)-N''-methylurea and methyl N'-(3-indolylmethyl)carbamate. The substrate specificity of BO suggests that selective inhibitors can be designed for the potential control of L. maculans.
authors.affiliation: Department of Chemistry, University of Saskatchewan, 110 Science Place, Saskatoon, SK, Canada. soledade.pedras@usask.ca
authors.forename: M Soledade C
authors.initials: MS
authors.lastname: Pedras
authors.forename: Mukund
authors.initials: M
authors.lastname: Jha
authors.forename: Zoran
authors.initials: Z
authors.lastname: Minic
authors.forename: Oladapo G
authors.initials: OG
authors.lastname: Okeola
mesh_terms: D000935:Antifungal Agents / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D001203:Ascomycota / Q000187:drug effects* / Q000378:metabolism / Q000472:pathogenicity; D019607:Brassicaceae / Q000378:metabolism / Q000382:microbiology; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D008658:Inactivation, Metabolic; D007211:Indoles / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D010935:Plant Diseases / Q000382:microbiology; D010936:Plant Extracts / Q000378:metabolism*; D013237:Stereoisomerism; D013329:Structure-Activity Relationship; D013859:Thiocarbamates / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D000094162:Phytoalexins
chemical_list: D000935:Antifungal Agents; D007211:Indoles; D010936:Plant Extracts; D013859:Thiocarbamates; C089020:brassinin; D000094162:Phytoalexins