Purification and characterization of platelet aggregation inhibitor from the venom of Bitis arietans
Тип публікації :
Стаття
Дата випуску :
2022
Автор(и) :
O. Platonov
V. Nikulina
Y. Kucheryavyi
V. Gryshchuk
Y. Stohniy
V. Chernyshenko
O. Slominskyi
A. Rebriev
K. Savchenko
Мова основного тексту :
Англійська
eKNUTSHIR URL :
Журнал :
Ukraine Biochemical Journal
Том :
94
Випуск :
5
ISSN :
2409-4943
Початкова сторінка :
7
Кінцева сторінка :
17
Цитування :
[APA 7] O., P., V., N., Y., K., V., G., Y., S., V., C., O., S., A., R., K., S., & Гарманчук, Л. В. (2022). Purification and characterization of platelet aggregation inhibitor from the venom of Bitis arietans. Ukraine Biochemical Journal, 94(5), 7–17. https://doi.org/10.15407/ubj94.05.007
[ДСТУ] Purification and characterization of platelet aggregation inhibitor from the venom of Bitis arietans / P. O. et al. Ukraine Biochemical Journal. 2022. Vol. 94, no. 5. P. 7—17. DOI: 10.15407/ubj94.05.007 (date of access: 18.07.2026).
Disintegrins are the antagonists of integrin receptors that can be found mostly in snakes’ venom. They can inhibit platelet aggregation, thus preventing the formation of blood clots. By blocking the integrin receptors of cancer cells, disintegrins can inhibit proliferation and metastasis. Thus, the search for new sources of disintegrins and development of methods of their purification is an important task of modern biotechnology. This work was dedicated to the purification and characterization of inhibiting polypeptides from Bitis arietans venom. Crude venom of B. arietans was fractionated using ion-exchange chromatography on Q Sepharose followed by size-exclusion chromatography on Superdex 75 using FPLC method. Analysis of molecular weight of protein components was performed using SDS-PAGE and MALDI-TOF analysis on Voyager-DE. Aggregation of platelet-rich plasma (PRP) in the presence of platelet aggregation inhibitor was investigated using aggregometry on the AR2110. MTT test was used for measuring HeLa cells proliferation and survival in vitro. Two-step chromatography allowed us to obtain fraction that contained polypeptides possessing the dose-dependent inhibitory action on adenosine diphosphate (ADP)-induced platelet aggregation in PRP. SDS-PAGE showed that obtained fraction contained two polypeptides with molecular weight 9.0 and 13.67 kDa according to MALDI-TOF analysis. Purified polypeptides inhibited ADP-induced platelet aggregation with IC50 0.09 mg/ml. However, 0.005 mg/ml of fraction suppressed viability of HeLa cells according to MTT test on 20%. Discovered biological effects of fractions allowed us to conclude the possible use of these polypeptides as anti-aggregatory or anti-proliferative agents. Keywords: antithrombotic action, disintegrins, glycoprotein IIb/IIIa, platelets, snake venom
Галузі знань та спеціальності :
09 Біологія
Галузі науки і техніки (FOS) :
Природничі науки
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Формат :
Adobe PDF
Розмір :
1.39 MB
Контрольна сума :
(MD5):af2a0bb907437979f42f3fe186375bd9
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