Purification, cloning and characterization of fragaceatoxin C, a novel actinoporin from the sea anemone Actinia fragacea

Bellomio, Augusto; Morante, Koldo; Barlič, Ariana; Gutiérrez-Aguirre, Ion; Viguera, Ana Rosa; González-Mañas, Juan Manuel

doi:10.1016/j.toxicon.2009.06.022

Cited by 60 publications

(74 citation statements)

References 57 publications

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“…The column volume V c was 21.2 ml as determined with acetone. The elution volume of monomeric FraC was V m ϭ 27.9 ml, higher than the column volume because of the unspecific interaction of the monomeric toxin with the column matrix (16).…”

Section: Methodsmentioning

confidence: 98%

“…Activity Measurements-Hemolytic activity was performed as described previously (16). Briefly, sheep RBC were washed in phosphate buffer and standardized at A 700 of 0.6.…”

Section: Methodsmentioning

confidence: 99%

“…This unique family of eukaryotic PFTs displays a high degree of sequence identity (16). Their globular structure presents a common fold based on a ␤-sandwich core flanked by two ␣-helices (17,18).…”

mentioning

confidence: 99%

“…Their globular structure presents a common fold based on a ␤-sandwich core flanked by two ␣-helices (17,18). The best characterized actinoporins are equinatoxin II (19), sticholysin II (20), and FraC (16). Actinoporins have been studied as potential immunotoxins (21) and serve as models to study protein-membrane interactions, a process of great relevance in neurodegenerative diseases (22,23).…”

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confidence: 99%

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A Pore-Forming Toxin Requires a Specific Residue for Its Activity in Membranes with Particular Physicochemical Properties

Morante

Caaveiro

Tanaka

et al. 2015

Journal of Biological Chemistry

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Background: It is unclear how actinoporins adapt to the broad range of physicochemical landscapes of biological membranes. Results: A single mutation in fragaceatoxin C (an actinoporin) prevents the formation of lytic pores in membranes in the liquid-ordered phase. Conclusion: Phe-16 is critical for pore formation of fragaceatoxin C in cholesterol-rich membranes. Significance: Site-directed mutagenesis generates lipid phase-sensitive pore-forming proteins.

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Section: Methodsmentioning

confidence: 98%

“…Activity Measurements-Hemolytic activity was performed as described previously (16). Briefly, sheep RBC were washed in phosphate buffer and standardized at A 700 of 0.6.…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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A Pore-Forming Toxin Requires a Specific Residue for Its Activity in Membranes with Particular Physicochemical Properties

Morante

Caaveiro

Tanaka

et al. 2015

Journal of Biological Chemistry

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“…In the second method, which is the more employed of the two, the area is kept constant and the increase in π that is produced by injecting the protein into the subphase is measured, as a function of time, until equilibrium is reached (Brockman 1999;Maget-Dana 1999). Monolayers have been used to study the membrane binding of diverse families of proteins, such as lipolytic toxins (Maget-Dana 1999), some cell transcription factors, myelin protein components (Maggio et al 2008), sphingomyelinases (Fanani et al 2010) and several members of the actinoporin family (Barlic et al 2004;Bellomio et al 2009) including Sts (Martinez et al 2007;Pedrera et al 2014Pedrera et al , 2015. On the other hand, liposomal vesicles are the most accepted systems as membrane models due to their bilayer organization and vesicular geometry.…”

Section: Understanding the Interaction Of Actinoporins With Membranesmentioning

confidence: 99%

Biophysical and biochemical strategies to understand membrane binding and pore formation by sticholysins, pore-forming proteins from a sea anemone

et al. 2017

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Actinoporins constitute a unique class of poreforming toxins found in sea anemones that are able to bind and oligomerize in membranes, leading to cell swelling, impairment of ionic gradients and, eventually, to cell death. In this review we summarize the knowledge generated from the combination of biochemical and biophysical approaches to the study of sticholysins I and II (Sts, StI/II), two actinoporins largely characterized by the Center of Protein Studies at the University of Havana during the last 20 years. These approaches include strategies for understanding the toxin structure-function relationship, the protein-membrane association process leading to pore formation and the interaction of toxin with cells. The rational combination of experimental and theoretical tools have allowed unraveling, at least partially, of the complex mechanisms involved in toxin-membrane interaction and of the molecular pathways triggered upon this interaction. The study of actinoporins is important not only to gain an understanding of their biological roles in anemone venom but also to investigate basic molecular mechanisms of protein insertion into membranes, protein-lipid interactions and the modulation of protein conformation by lipid binding. A deeper knowledge of the basic molecular mechanisms involved in Sts-cell interaction, as described in this review, will support the current investigations conducted by our group which focus on the design of immunotoxins against tumor cells and antigen-releasing systems to cell cytosol as Stsbased vaccine platforms.

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Sticholysin I–II oligomerization in the absence of membranes

et al. 2022

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Sticholysins are pore-forming toxins produced by the sea anemone Stichodactyla helianthus. When they encounter a sphingomyelin-containing membrane, these proteins bind to it and oligomerize, a process that ends in pore formation. Mounting evidence indicates that StnII can favour the activity of StnI. Previous results have shown that these two isotoxins can oligomerize together. Furthermore, StnII appeared to potentiate the activity of StnI through the membrane-binding step of the process. Hence, isotoxin interaction should occur prior to membrane encounter. Here, we have used analytical ultracentrifugation to investigate the oligomerization of Stns in solution, both separately and together. Our results indicate that while StnI seems to be more prone to oligomerize in water solution than StnII, a small percentage of StnII in StnI-StnII mixtures promotes oligomerization.

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Purification, cloning and characterization of fragaceatoxin C, a novel actinoporin from the sea anemone Actinia fragacea

Cited by 60 publications

References 57 publications

A Pore-Forming Toxin Requires a Specific Residue for Its Activity in Membranes with Particular Physicochemical Properties

A Pore-Forming Toxin Requires a Specific Residue for Its Activity in Membranes with Particular Physicochemical Properties

Biophysical and biochemical strategies to understand membrane binding and pore formation by sticholysins, pore-forming proteins from a sea anemone

Sticholysin I–II oligomerization in the absence of membranes

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