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Mar Drugs. 2015 Dec 30;14(1):7. doi: 10.3390/md14010007.

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

Author information

1
Faculty of Pharmacy, Meijo University, Tempaku, Nagoya 468-8503, Japan. arojanas@riem.nagoya-u.ac.jp.
2
Department of Brain Functions, Division of Stress Adaptation and Protection, Research Institute of Environmental Medicine, Nagoya University, Nagoya 464-8601, Japan. arojanas@riem.nagoya-u.ac.jp.
3
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan. g0973345@ccalumni.meijo-u.ac.jp.
4
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan. 100973149@ccalumni.meijo-u.ac.jp.
5
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan. 110973437@ccalumni.meijo-u.ac.jp.
6
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan. asukabe@bj8.so-net.ne.jp.
7
Faculty of Pharmacy, Meijo University, Tempaku, Nagoya 468-8503, Japan. kiharada@meijo-u.ac.jp.

Abstract

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.

KEYWORDS:

aeruginosins; anticoagulant; blood coagulation cascade; cyanobacteria; fVIIa-sTF inhibitors; peptides; toxic Microcystis

PMID:
26729138
PMCID:
PMC4728504
DOI:
10.3390/md14010007
[Indexed for MEDLINE]
Free PMC Article

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