Purification, characterization and immobilization of proteinase inhibitors from Stichodactyla helianthus

Delfín, Julieta; Martínez, Isabel Lorente; Antuch, Walfrido; Morera, Vivian; González, Yoel Martínez; Rodrı́guez, Rolando; Márquez, Margarita; Saroyán, Angélika; Ларионова, Н. И.; Díaz, Javier Santovenia; Padrón, Gabrìel; Chávez, María Á.

doi:10.1016/s0041-0101(96)00114-6

Cited by 78 publications

(59 citation statements)

References 19 publications

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“…Our results displayed that ShPI-I was a more potent inhibitor for Leishmania serine protease activities and promastigote survival than TPCK or Bza. As previously mentioned, although ShPI-I also inhibits other proteases, it is much more specific and effective for serine proteases (Delfin et al 1996). Therefore, the action of this inhibitor might quite possibly be due to an accumulative inhibition of other protease activity, although the molar concentration of ShPI-I applied to inhibit cysteine or aspartic protease is much higher than the amount employed in the present study.…”

Section: Discussionmentioning

confidence: 64%

“…As expected, ShPI-I in lower molar concentration (10 −6 and 10 −5 M) than TPCK and Bza was more effective in reducing promastigote viability. This peptide is an irreversible, tight-binding protease inhibitor and is effective in very low molar concentrations (Delfin et al 1996). Although the effects of other molecular from marine organisms on the growth of Leishmania promastigotes have already been studied (Savoia et al 2004), this is the first study that reports the effects of protease inhibitor from marine organisms on Leishmania proteases and how this inhibition may affect the viability of this protozoa.…”

Section: Discussionmentioning

confidence: 98%

“…Inhibition was expressed as the percentage of the appropriate control activity (100%). The Kunitz type inhibitor was purified from sea anemone Stichodactyla helianthus as described previously (Delfin et al 1996).…”

Section: Effect Of Shpi-i On Enzymatic Activitymentioning

confidence: 99%

“…We have previously reported that Bza completely inhibited the LSPI activity (SilvaLopez and Giovanni De Simone 2004a) and 80% LSPIII activity (Silva-Lopez et al 2005) but did not affect the LSPII (Silva-Lopez and Giovanni De Simone 2004b), while TPCK inhibited all enzymes in different degrees, TLCK had a moderate influence on the LSP II and LSP III activity but did not affect the detergent-soluble Leishmania serine protease (Silva-Lopez and Giovanni De Simone 2004a, b;Silva-Lopez et al 2005). Furthermore, we also analyzed the effect of a high molecular weight inhibitor from sea anemone Stichodactyla helianthus (ShPI-I), which is a Kunitz-type inhibitor of 6,110.6 Da that exhibits a broad specificity for serine, cysteine, and aspartic proteases (Delfin et al 1996). Regardless of the fact that ShPI-I inhibits other peptidases, it was demonstrated that it has a great specificity for serine proteases due to the lowest Ki values obtained for serine in comparison to the Ki of cysteine and aspartic proteases (Delfin et al 1996).…”

Section: Introductionmentioning

confidence: 99%

“…Furthermore, we also analyzed the effect of a high molecular weight inhibitor from sea anemone Stichodactyla helianthus (ShPI-I), which is a Kunitz-type inhibitor of 6,110.6 Da that exhibits a broad specificity for serine, cysteine, and aspartic proteases (Delfin et al 1996). Regardless of the fact that ShPI-I inhibits other peptidases, it was demonstrated that it has a great specificity for serine proteases due to the lowest Ki values obtained for serine in comparison to the Ki of cysteine and aspartic proteases (Delfin et al 1996). Furthermore, the higher specificity of ShPI-I observed for serine proteases might also be explained by the fact that this inhibitor demonstrated a nearly identical molecular architecture to the bovine pancreatic trypsin inhibitor (BPTI), an important mammalian serine protease inhibitor.…”

Section: Introductionmentioning

confidence: 99%

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Effects of serine protease inhibitors on viability and morphology of Leishmania (Leishmania) amazonensis promastigotes

et al. 2007

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To investigate the importance of serine proteases in Leishmania amazonensis promastigotes, we analyzed the effects of classical serine protease inhibitors and a Kunitz-type inhibitor, obtained from sea anemone Stichodactyla helianthus (ShPI-I), on the viability and morphology of parasites in culture. Classical inhibitors were selected on the basis of their ability to inhibit L. amazonensis serine proteases, previously described. The N-tosyl-L: -phenylalanine chloromethyl ketone (TPCK) and benzamidine (Bza) inhibitors, which are potential Leishmania proteases inhibitors, in all experimental conditions reduced the parasite viability, with regard to time dependence. On the other hand, N-tosyl-lysine chloromethyl ketone (TLCK) did not significantly affect the parasite viability, as it was poor Leishmania enzymes inhibitor. Ultrastructural analysis demonstrated that both Bza and TPCK induced changes in the flagellar pocket region with membrane alteration, including bleb formation. However, TPCK effects were more pronounced than those of Bza in Leishmania flagellar pocket in plasma membrane, and intracellular vesicular bodies was visualized. ShPI-I proved to be a powerful inhibitor of L. amazonensis serine proteases and the parasite viability. The ultrastructural alterations caused by ShPI-I were more dramatic than those induced by the classical inhibitors. Vesiculation of the flagellar pocket membrane, the appearance of a cytoplasmic vesicle that resembles an autophagic vacuole, and alterations of promastigotes shape resulted.

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 98%

Section: Effect Of Shpi-i On Enzymatic Activitymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Effects of serine protease inhibitors on viability and morphology of Leishmania (Leishmania) amazonensis promastigotes

et al. 2007

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Cnidaria and Ctenophora–2

Kornprobst

2014

Encyclopedia of Marine Natural Products

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The article contains sections titled: Membrane Constituents and Secondary Metabolites of Hexacorallia (Anthozoa) Fatty Acids, Oxylipins, and Acetylenic Derivatives Sterols and Ecdysteroids Terpenes Carotenoids of Sea Anemones Aromatic Derivatives Ceramides and Lipidic α‐Amino Acids Examples of Nitrogenous Pigments: Zoanthoxanthins and Calliactine Palytoxins ( PTXs ) Mycosporines Betaines, Phosphobetaines, Purines, and Other Nitrogen‐Containing Derivatives Zoanthamine Alkaloids Cytolysins, Neuropeptides, and Other Venoms of Sea Anemones Venoms and Other Metabolites of Medusozoa Some Data on the Cubozoa and Other Jellyfish Bioluminescence of Jellyfish and Other Cnidaria Some Results on the Secondary Metabolites of Other Hydrozoa Ctenophora

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Immunohistochemical targeting of sea anemone cytolysins on tentacles, mesenteric filaments and isolated nematocysts ofStichodactyla helianthus

et al. 2006

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The toxicity of biomolecules obtained from sea anemones in vitro does not necessarily justify their function as toxins in the physiology of the anemone. That is why anatomical and physiological considerations must be taken into account in order to define their physiological role in the organism. In this work, antibodies generated to Sticholysin II, a cytolysin produced by the Caribbean Sea anemone Stichodactyla helianthus, are used as specific markers to explore the sites of production and storage of the cytolysin in the sea anemone. The immunoperoxidase staining developed gave specific dark-brown staining in tentacles and mesenteric filaments as well as in basitrichous nematocysts isolated from tentacles of S. helianthus. These results support the role of these proteins as toxins in the physiology of the anemone, especially in functions such as in predation, defense and digestion.

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Purification, characterization and immobilization of proteinase inhibitors from Stichodactyla helianthus

Cited by 78 publications

References 19 publications

Effects of serine protease inhibitors on viability and morphology of Leishmania (Leishmania) amazonensis promastigotes

Effects of serine protease inhibitors on viability and morphology of Leishmania (Leishmania) amazonensis promastigotes

Cnidaria and Ctenophora–2

Immunohistochemical targeting of sea anemone cytolysins on tentacles, mesenteric filaments and isolated nematocysts ofStichodactyla helianthus

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