Comprehensive study of proteins that interact with microcystin-LR

作者：Mori Tomoko^*; Kubo Takuya; Kaya Kunimitsu; Hosoya Ken

来源：Analytical and Bioanalytical Chemistry, 2012, 402(3): 1137-1147.

DOI：10.1007/s00216-011-5514-1

• Microcystin-LR
• Target proteins
• Affinity resin
• Nano-LC/MS/MS

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.

• 出版日期2012-1