• BMB Reports
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• 제40권5호
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• pp.832-838
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• 2007

A novel inhibitory protein against blood coagulation factor Va (FVa) was purified from muscle protein of granulated ark (Tegillarca granosa, order Arcoida, marine bivalvia) by consecutive FPLC method using anion exchange and gel permeation chromatography. In the results of ESI-QTOF tandem mass analysis and database research, it was revealed that the purified T. granosa anticoagulant protein (TGAP) has 7.7 kDa of molecular mass and its partial sequence, HTHLQRAPHPNALGYHGK, has a high identity (64%) with serine/threonine kinase derived from Rhodopirellula baltica (order Planctomycetales, marine bacteria). TGAP could potently prolong thrombin time (TT), corresponding to inhibition of thrombin (FIIa) formation. Specific factor inhibitory assay showed that TGAP inhibits FVa among the major components of prothrombinase complex. In vitro assay for direct-binding affinity using surface plasmon resonance (SPR) spectrometer indicated that TGAP could be directly bound with FVa. In addition, the binding affinity of FVa to FII was decreased by addition of TGAP in dose-dependant manner ($IC_{50}$ value = 77.9 nM). These results illustrated that TGAP might interact with a heavy chain of FVa ($FVa_H$) bound to FII in prothrombin complex. The present study elucidated that non-cytotoxic T. granosa anticoagulant protein (TGAP) bound to FVa can prolong blood coagulation time by inhibiting conversion of FII to FIIa in blood coagulation cascade. In addition, TGAP did not significantly (P < 0.05) show fibrinolytic activity and cytotoxicity on venous endothelial cell line (ECV 304).

• 대한한방내과학회지
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• 제27권3호
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• pp.589-602
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• 2006

Hwangryunagyotang is supposed to have significant effects on some sorts of cardiovascular diseases like atherosclerosis. For this study. ACAT inhibitor was put in LDLR -/- mice to derive free cholesterol from it. This was to examine the effectiveness of Hwangryunnagyotang on its protecting and recovering function with endothelial cells damaged by free cholesterol through experimental. The results reported below. Hwangryunagyotang suppressed the crystallization of reactive oxygen species in macrophages and the numbers of free cholesterol crystal plate structured and reduced fragmentation of nucleus in ECV 304 cell strain by ACAT inhibitor significantly. Hwangryunagyotang also suppressed the necrosis of tissue in LDLR -/- mice' (treated with ACAT inhibitor) inflammatory portion which is adjacent to aortic root, proximal aorta and carotid artery by immunohistochemistry and fluorescence microscopy. On the whole, Hwangryunagyotang suppressed the necrosis of endothelial cells and especially it's effcet for the necrosis of para-myocardial tissues by free cholesterol. With this result, I suggest Hwangryunagyotang might have protective and recovery effects on atherosclerosis, so we need to carry on this study henceforth clinically and experimentally as well.