Highly stable glycosylated serine protease from the medicinal plant Euphorbia milii

Yadav, Subhash Chandra; Pande, Monu; Jagannadham, Medicherla V.

doi:10.1016/j.phytochem.2006.06.002

Cited by 80 publications

(71 citation statements)

References 53 publications

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“…5 and 6) on fibrinolytic activity and stability of StSBTc-3 were similar with data reported for other plant; bacterial and green algae serine proteases with fibrinogenolytic activity [1,14,16,38,40,46]. StBTc-3 was found to be active at pH values ranging from 7 to 11 and temperatures from 20 C to 60 C. The optimal pH for the fibrinogenolytic activity was 8 ( Fig.…”

Section: Effect Of Ph and Temperature On The Fibrinogenolytic Activitsupporting

confidence: 86%

“…A comparable cleavage pattern was observed in the course of insulin b-chain hydrolysis with plant serine proteases from Cucumis melo, Trichosanthes cucumeroides, Parthenium agentatum, Maclura pomifera (named as Macluralisin). As reported for a serine protease isolated from Helianthus annuus, a fungal proteinase A1 from Trichosanthes cucumeroides and human plasmin [40,41], StSBTc-3 hydrolyzes the amino acid bond between Arg22-Gly23 (Fig. 4).…”

Section: Digestion Of Oxidized Insulin B-chain By Stsbtc-3mentioning

confidence: 68%

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Fibrin(ogen)olytic and antiplatelet activities of a subtilisin-like protease from Solanum tuberosum (StSBTc-3)

et al. 2016

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a b s t r a c tPlant serine proteases have been widely used in food science and technology as well as in medicine. In this sense, several plant serine proteases have been proposed as potential anti-coagulants and antiplatelet agents. Previously, we have reported the purification and identification of a plant serine protease from Solanum tuberosum leaves. This potato enzyme, named as StSBTc-3, has a molecular weight of 72 kDa and it was characterized as a subtilisin like protease. In this work we determine and characterize the biochemical and medicinal properties of StSBTc-3. Results obtained show that, like the reported to other plant serine proteases, StSBTc-3 is able to degrade all chains of human fibrinogen and to produces fibrin clot lysis in a dose dependent manner. The enzyme efficiently hydrolyzes b subunit followed by partially hydrolyzed a and g subunits of human fibrinogen. Assays performed to determine StSBTc-3 substrate specificity using oxidized insulin b-chain as substrate, show seven cleavage sites: Asn3-Gln4; Cys7-Gly8; Glu13-Ala14; Leu15-Tyr16; Tyr16-Leu17; Arg22-Gly23 and Phe25-Tyr26, all of them were previously reported for other serine proteases with fibrinogenolytic activity. The maximum StSBTc-3 fibrinogenolytic activity was determined at pH 8.0 and at 37 C. Additionally, we demonstrate that StSBTc-3 is able to inhibit platelet aggregation and is unable to exert cytotoxic activity on human erythrocytes in vitro at all concentrations assayed. These results suggest that StSBTc-3 could be evaluated as a new agent to be used in the treatment of thromboembolic disorders such as strokes, pulmonary embolism and deep vein thrombosis.

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Section: Effect Of Ph and Temperature On The Fibrinogenolytic Activitsupporting

confidence: 86%

Section: Digestion Of Oxidized Insulin B-chain By Stsbtc-3mentioning

confidence: 68%

Fibrin(ogen)olytic and antiplatelet activities of a subtilisin-like protease from Solanum tuberosum (StSBTc-3)

et al. 2016

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“…6). Human fibrinogen is a heterotrimeric protein composed of Aα, Bβ, and γ [5]. Each subunit (Aα, Bβ, and γ) has an estimated molecular weight of 66, 55, and 48 kDa, respectively [21].…”

Section: Resultsmentioning

confidence: 99%

The Effects of Metal Ions in Euphorbia cf. lactea Latex on the Fibrinogenolytic Activity of a Plant Protease

Siritapetawee

Limphirat

Kantachot

et al. 2014

Appl Biochem Biotechnol

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Two synchrotron-based techniques, synchrotron X-ray fluorescence (SXRF) and X-ray absorption spectroscopy (XAS), have demonstrated that Ca(2+) and Zn(2+) were the major metal ions distributed in the natural latex of Euphorbia cf. lactea. Both metal ions were found to affect the fibrinogenolytic activity of a homodimeric protease purified from the latex of this plant. The dimeric protein had an estimated molecular mass of about 82 kDa analyzed by SDS-PAGE. Therefore this protein was called as EuP-82. Based on the results of circular dichroism (CD) spectroscopy and the fibrinolytic activity measurement, it was found that Ca(2+) could activate the proteolytic activity of the enzyme by stabilizing its backbone structure. The intact conformation of EuP-82 was predicted from CD spectrum, which consisted of 51 % α-helix and 9 % β-sheet. Zn(2+) (10 mM) could decrease the fibrinolytic activity of EuP-82 to 30 ± 1 %. CD spectrum also supported that the inhibitory effect of Zn(2+) on the enzyme activity occurred by the drastic change of the enzyme structure with increasing the random coil conformation and by switching between α-helix and β-sheet structure. These results could be of first importance for further application to use EuP-82, the natural source protease as a potential drug for the thrombosis treatment. The fibrinolytic activity of EuP-82 may be enhanced by plasma Ca(2+) which generally involves in human hemostasis system.

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“…However, serine proteases are found in the latex of Euphorbiaceae. This type of enzyme was obtained, for example, from the latex of Euphorbia supina (Arima et al, 2000) and Euphorbia milii, with potential applications in food industry (Yadav, Pande, Jagannadham, 2006;Moro et al, 2008;Fonseca et al, 2010). Also from Euphorbia hirta, a serine protease, with fibrinolytic activity and potential industrial and therapeutic applications, was purified (Patel, Kawale, Sharma, 2012).…”

Section: Resultsmentioning

confidence: 99%

Proteinase activity in latex of three plants of the family Euphorbiaceae

Sobottka

Tonial

Sytwala

et al. 2014

Braz. J. Pharm. Sci.

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In the family of Euphorbiaceae, the genera Euphorbia and Sapium are known to contain essentially latex-bearing species. In the present study, the latex of Euphorbia selloi (Klotzsch & Garcke) Boiss., Euphorbia papillosa A.St.-Hil., and Sapium glandulosum (L.) Morong, plants native from Brazil, were examined concerning proteolytic activity. All studied species have proteins with significant proteolytic activity and E. papillosa has the greatest specific activity. Aiming to verify the type of protease present, an assay with different inhibitors was performed. In the three tested plants, the proteolytic activity was significantly inhibited by a serine protease inhibitor 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride (AEBSF). Using techniques of electrophoresis with polyacrylamide gels (SDS-PAGE), the subunits of proteins were separated according to their molecular masses, and the protein activity was visually detected by zymography.Uniterms: Euphorbiaceae/species/phytochemistry. Euphorbia papillosa/phytochemistry. Euphorbia selloi/ phytochemistry. Sapium glandulosum/phytochemistry. Euphorbia papillosa/proteinase activity. Euphorbia selloi/proteinase activity. Sapium glandulosum/proteinase activity. Endopeptidase. Gel electrophoresis.Dentro da família Euphorbiaceae, os gêneros Euphorbia e Sapium são conhecidos por incluírem basicamente espécies produtoras de látex. No presente estudo, o látex das plantas Euphorbia selloi (Klotzsch & Garcke) Boiss., Euphorbia papillosa A.St.-Hil. e Sapium glandulosum (L.) Morong, espécies nativas do Brasil, foi analisado em relação à atividade proteolítica. Todas as amostras analisadas possuem proteínas com significativa atividade, sendo que o látex da espécie E. papillosa apresenta a maior atividade específica. Com o objetivo de analisar quais os tipos de proteases responsáveis pela atividade proteolítica, realizaram-se ensaios com diferentes inibidores. Nas três plantas testadas a atividade foi inibida significativamente pelo cloridrato de 4-(fluoreto de 2-aminoetilbenzenossulfonil) (AEBSF), um inibidor de serino-proteases. Utilizando técnicas de eletroforese em gel de poliacrilamida (SDS-PAGE), as subunidades das proteínas foram separadas de acordo com sua massa molecular e, através da zimografia, a atividade proteolítica pode ser detectada visualmente. Unitermos

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Highly stable glycosylated serine protease from the medicinal plant Euphorbia milii

Cited by 80 publications

References 53 publications

Fibrin(ogen)olytic and antiplatelet activities of a subtilisin-like protease from Solanum tuberosum (StSBTc-3)

Fibrin(ogen)olytic and antiplatelet activities of a subtilisin-like protease from Solanum tuberosum (StSBTc-3)

The Effects of Metal Ions in Euphorbia cf. lactea Latex on the Fibrinogenolytic Activity of a Plant Protease

Proteinase activity in latex of three plants of the family Euphorbiaceae

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