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Open AccessArticle

Determination of FVIIa-sTF Inhibitors in Toxic Microcystis Cyanobacteria by LC-MS Technique

1
Faculty of Pharmacy, Meijo University, Tempaku, Nagoya 468-8503, Japan
2
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan
3
Department of Brain Functions, Division of Stress Adaptation and Protection, Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-8601, Japan
*
Authors to whom correspondence should be addressed.
Academic Editor: Vitor H. Pomin
Mar. Drugs 2016, 14(1), 7; https://doi.org/10.3390/md14010007
Received: 25 September 2015 / Revised: 18 December 2015 / Accepted: 21 December 2015 / Published: 30 December 2015
(This article belongs to the Special Issue Marine Anticoagulants and Antithrombotics)
The blood coagulation cascade involves the human coagulation factors thrombin and an activated factor VII (fVIIa). Thrombin and fVIIa are vitamin-K-dependent clotting factors associated with bleeding, bleeding complications and disorders. Thrombin and fVIIa cause excessive bleeding when treated with vitamin-K antagonists. In this research, we explored different strains of toxic Microcystis aeruginosa and cyanobacteria blooms for the probable fVIIa-soluble Tissue Factor (fVIIa-sTF) inhibitors. The algal cells were subjected to acidification, and reverse phase (ODS) chromatography-solid phase extraction eluted by water to 100% MeOH with 20%-MeOH increments except for M. aeruginosa NIES-89, from the National Institute for Environmental Studies (NIES), which was eluted with 5%-MeOH increments as an isolation procedure to separate aeruginosins 89A and B from co-eluting microcystins. The 40%–80% MeOH fractions of the cyanobacterial extract are active against fVIIa-sTF. The fVIIa-sTF active fractions from cultured cyanobacteria and cyanobacteria blooms were subjected to liquid chromatography-mass spectrometry (LC-MS). The 60% MeOH fraction of M. aeruginosa K139 exhibited an m/z 603 [M + H]+ attributed to aeruginosin K139, and the 40% MeOH fraction of M. aeruginosa NIES-89 displayed ions with m/z 617 [M − SO3 + H]+ and m/z [M + H]+ 717, which attributed to aeruginosin 89. Aeruginosins 102A/B and 298A/B were also observed from other toxic strains of M. aeruginosa with positive fVIIa-sTF inhibitory activity. The active fractions contained cyanobacterial peptides of the aeruginosin class as fVIIa-sTF inhibitors detected by LC-MS. View Full-Text
Keywords: cyanobacteria; toxic Microcystis; anticoagulant; fVIIa-sTF inhibitors; peptides; aeruginosins; blood coagulation cascade cyanobacteria; toxic Microcystis; anticoagulant; fVIIa-sTF inhibitors; peptides; aeruginosins; blood coagulation cascade
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Anas, A.R.J.; Nakajima, A.; Naruse, C.; Tone, M.; Asukabe, H.; Harada, K.-I. Determination of FVIIa-sTF Inhibitors in Toxic Microcystis Cyanobacteria by LC-MS Technique. Mar. Drugs 2016, 14, 7.

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