Isolation and Characterization of a 32-kDa Fibrinolytic Enzyme (FE-32kDa) from Gloydius blomhoffii siniticus Venom: Fibrinolytic Enzyme from Gloydius blomhoffii siniticus Venom

Journal of Pharmacopuncture
Joung-Yoon KimSuk-Ho Choi

Abstract

This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as Fe(2+) and Hg(2+) inhibited the fibrin hydrolysis completely, but Zn(2+) enhanced it. FE-32kDa hydrolyzed α-chain but did not hydrolyze β-chain and γ-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydro...Continue Reading

References

Nov 15, 1989·Archives of Biochemistry and Biophysics·E N BaramovaJ W Fox
Dec 5, 1998·Toxicon : Official Journal of the International Society on Toxinology·F S Markland
Oct 1, 1952·Archives of Biochemistry and Biophysics·T ASTRUP, S MULLERTZ
May 11, 2005·Toxicon : Official Journal of the International Society on Toxinology·S Swenson, F S Markland
Apr 1, 2010·Toxins·Francis S Markland, Steve Swenson

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Citations

Aug 5, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Etsuko Oyama, Hidenobu Takahashi
Dec 4, 2019·Journal of Rural Medicine : JRM·Ikuto TakeuchiYouichi Yanagawa

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Methods Mentioned

BETA
electrophoresis
Protein Assay
gel filtration
ion exchange chromatography

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