Izvorni znanstveni članak
https://doi.org/10.3325/cmj.2017.58.118
Antiplatelet and antiproliferative action of disintegrin from Echis multisquamatis snake venom
Volodymyr Chernyshenko
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Natalia Petruk
; Educational and Scientific Centre “Institute of Biology” Taras Shevchenko National University, Kyiv, Ukraine
Darya Korolova
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Ludmila Kasatkina
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Olha Gornytska
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Tetyana Platonova
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Tamara Chernyshenko
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Andriy Rebriev
; Protein Structure and Functions Department, Palladin Institute of biochemistry NAS of Ukraine, Kyiv, Ukraine
Olena Dzhus
; Educational and Scientific Centre “Institute of Biology” Taras Shevchenko National University Kyiv, Ukraine
Liudmyla Garmanchuk
; Educational and Scientific Centre “Institute of Biology” Taras Shevchenko National University Kyiv, Ukraine
Eduard Lugovskoy
; Educational and Scientific Centre “Institute of Biology” Taras Shevchenko National University Kyiv, Ukraine
Sažetak
Aim To purify the platelet aggregation inhibitor from Echis
multisquamatis snake venom (PAIEM) and characterize its
effect on platelet aggregation and HeLa cell proliferation.
Methods Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
(SDS-PAGE) and matrix assisted laser desorption/
ionization time-of-flight (MALDI-TOF) were used for
PAIEM identification. Platelet aggregation in the presence
of PAIEM was studied on aggregometer Solar-AP2110. The
changes of shape and granularity of platelets in the presence
of PAIEM were studied on flow cytometer COULTER
EPICS XL, and degranulation of platelets was estimated
using spectrofluorimetry. Indirect enzyme-linked immunosorbent
assay was used for the determination of target
of PAIEM on platelet surface. An assay based on 3-(4,5-dimethylthiazol-
2-yl)-2,5-diphenyltetrazolium bromide was
used to evaluate the effect of PAIEM on the proliferation of
HeLa cells in cell culture.
Results The molecular weight of the protein purified from
Echis multisquamatis venom was 14.9 kDa. Half-maximal inhibitory
concentration (IC50) of PAIEM needed to inhibit adenosine
diphosphate (ADP)-induced platelet aggregation
was 7 μM. PAIEM did not affect thrombin- or ADP-induced
platelet activation, but it did prevent binding of the anti-
IIb antibody to glycoprotein IIb/IIIa (GPIIbIIIa)-receptor of
adhered platelets and inhibited the viability of HeLa cells
by 54%.
Conclusion As a member of the disintegrin family, PAIEM
inhibited platelet aggregation and cell proliferation possibly
by blocking integrin-mediated interactions. However,
it did not impair cellular signaling causing any changes in
platelet shape and granularity and did not affect ADP-induced
platelet degranulation. This disintegrin was shown
to be a potent inhibitor of integrin-mediated cellular interactions
including platelet aggregation or cancer cell proliferation.
Ključne riječi
Hrčak ID:
181579
URI
Datum izdavanja:
15.4.2017.
Posjeta: 971 *