Cucumisin-like protease from the latex of Euphorbia supina

Arima, Kazunari; Uchikoba, Tetsuya; Yonezawa, Hiroo; Shimada, M.; Kaneda, Makoto

doi:10.1016/s0031-9422(99)00605-6

Cited by 50 publications

(29 citation statements)

References 21 publications

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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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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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“…The same peptide was also preferentially hydrolyzed by cucumisin and the subtilisin-like serine endopeptidases from fruits of Maclura pomifera (Rudenskaya et al 1995) and Melothria japonica (Uchikoba et al 2001), and from the latex of dandelion roots (taraxalisin; Rudenskaya et al 1998) and Euphorbia supina (Arima et al 2000). The K m for SEP-1 hydrolysis of the peptide AAPL pNA (2.6 mM) was relatively high, indicating it had an intermediate affinity for the substrate.…”

Section: Discussionmentioning

confidence: 98%

SEP-1 - a subtilisin-like serine endopeptidase from germinated seeds of Hordeum vulgare L. cv. Morex

Fontanini

Jones

2002

Planta

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Proteolysis is crucial for all living cells. It regulates protein processing, intracellular protein levels and removes abnormal or damaged proteins from the cell, working as a cellular housekeeper. By means of proteolysis, cells can control the short-lived regulatory proteins that affect processes such as signal transduction and reception, transcription, division and cellular growth. Proteolysis also furnishes amino acids for the de novo synthesis of proteins. In germinating seeds, its main role is to degrade storage proteins into small peptides and amino acids that can be used by the embryo during autotrophic growth. We have isolated and purified a serine endopeptidase, one of the many proteolytic enzymes that occur in germinated barley seeds (green malt), using chromatofocusing and DEAE-, CM-, and size-exclusion chromatographies. The enzyme, named SEP-1, has a molecular weight of 70 kDa, as estimated by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography. SEP-1 was detected and measured by its ability to digest gelatin in gels and to hydrolyze the synthetic substrate N-succinyl Ala-Ala-Pro-Leu p-nitroanilide. The hydrolysis of the synthetic substrate was optimal at pH 6.5 and 50 degrees C with a K(m) of 2.6 mM. The enzyme was inhibited by phenylmethylsulfonyl fluoride and p-amidinophenyl methanesulfonyl fluoride but not by any other class-specific inhibitor, suggesting it was a serine endopeptidase. Its amino acid sequence was similar to those of other plant subtilisin-like serine peptidases (EC 3.4.21), especially to the cucumisin-like group. SEP-1 was present in resting seeds, and its activity increased during germination in all of the malted barley tissues except for the endosperm, where it never occurred, suggesting that the enzyme is not likely involved in storage-protein degradation.

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“…and diff erent protein classes (Agrawal & Konno 2009). Various latexes are known to contain glycosidases (Giordani & Lafon 1993), proteases (Kimmel & Smith 1954;Sgarbieri et al 1964;Lynn & Clevette-Radford 1987b;Arribére et al 1998;Arima et al 2000;Tomar et al 2008), acid phosphatases (Lynn & Clevette-Radford 1987a), amylases (Lynn & Clevette-Radford 1987a), chitinases (Jekel et al 1991), hevein (Van Parijs et al 1991), proteinase inhibitors (Archer 1983;Lin & Lu 1994), b-1,3-glucanase (Chye & Cheung 1995) and various other enzymatic activities (Lynn & Clevette-Radford 1987b;Agrawal & Konno 2009).…”

Section: Discussionmentioning

confidence: 99%

Leaf structure, microanalysis and characterization of the latex protein profile of Pachystroma longifolium (Nees) I.M. Jonhst. (Euphorbiaceae) in a seasonally dry Atlantic Forest

et al. 2011

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ABSTRACT(Leaf structure, microanalysis and characterization of the latex protein profile of Pachystroma longifolium (Nees) I.M. Jonhst. (Euphorbiaceae) in a seasonally dry Atlantic Forest). Pachystroma longifolium is an evergreen species of Euphorbiaceae that occurs in tabuleiro forest, a type of lowland Atlantic Forest. The site chosen for this study was a fragment of tabuleiro forest that has a history of being selectively logged for timber. P. longifolium is very successful at colonizing disturbed areas in this forest fragment in comparison with preserved fragments. In the present work, which was based on using different microscopy (electron and light microscopy, X-ray analysis) and biochemistry techniques, we describe the leaf and latex traits of P. longifolium and their role as defense mechanisms against desiccation and herbivory. Our results suggest the richness of P. longifolium populations, in the disturbed forest area studied, is primarily a consequence of unpalatable leaves because of an abundance of calcium oxalate crystals in the subjacent epidermis; the presence of phenolic compounds in the adaxial surface; and a laticifer system that contains a complex mixture of proteins, which provides resistance to herbivores. In addition, this species is resistant to desiccation during dry periods because of an extensive amount of wax that occurs on the outer cell walls of the epidermis, and its ability to retain water because of a biseriate epidermis.

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Cucumisin-like protease from the latex of Euphorbia supina

Cited by 50 publications

References 21 publications

Proteinase activity in latex of three plants of the family Euphorbiaceae

Proteinase activity in latex of three plants of the family Euphorbiaceae

SEP-1 - a subtilisin-like serine endopeptidase from germinated seeds of Hordeum vulgare L. cv. Morex

Leaf structure, microanalysis and characterization of the latex protein profile of Pachystroma longifolium (Nees) I.M. Jonhst. (Euphorbiaceae) in a seasonally dry Atlantic Forest

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