title: Comprehensive study of proteins that interact with microcystin-LR.
abstract: We carried out a comprehensive study of proteins that exhibit specific interactions with a naturally occurring toxin, microcystin (MC)-LR, in order to gain insight into the unknown underlying mechanism of MC virulence. This audacious study employed a simple affinity test that used MC-LR immobilized on an original ethylene oxide based monolithic solid phase (Moli-gel), and swine liver lysate. Some of the proteins that interacted with MC-LR on this original affinity resin were separated by SDS-PAGE, measured by nano-LC/MS/MS after trypsin digestion, and identified using a Mascot database search. Protein sequence analyses revealed that glutathione S-transferase (GST) was one of the candidate target proteins for MC-LR. This protein was confirmed as a target protein for MC-LR based on the results of for the inhibition of an enzymatic reaction by Dhb-MC-LR. Moreover, L-3-hydroxyacyl coenzyme A dehydrogenase (HDHA) was shown to be one of the proteins that specifically interacts with MC-LR. Our results demonstrated that our analytical systems based on an original affinity resin and nano-LC/MS/MS were effective for target protein research.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, Aoba 6-6-20, Aramaki, Aoba, Sendai 980-8579, Japan. mori@fmp.kankyo.tohoku.ac.jp
authors.forename: Tomoko
authors.initials: T
authors.lastname: Mori
authors.forename: Takuya
authors.initials: T
authors.lastname: Kubo
authors.forename: Kunimitsu
authors.initials: K
authors.lastname: Kaya
authors.forename: Ken
authors.initials: K
authors.lastname: Hosoya
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism*; D002846:Chromatography, Affinity; D000458:Cyanobacteria / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D008387:Marine Toxins; D052998:Microcystins / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism*; D013552:Swine; D053719:Tandem Mass Spectrometry
chemical_list: D001427:Bacterial Toxins; D008387:Marine Toxins; D052998:Microcystins; D011506:Proteins; D005982:Glutathione Transferase; C057862:cyanoginosin LR