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dc.creatorAlvarado Mesén, Javier
dc.creatorSolano Campos, Frank
dc.creatorCanet, Liem
dc.creatorPedrera, Lohans
dc.creatorHervis, Yadira P.
dc.creatorSoto, Carmen
dc.creatorBorbón, Henry
dc.creatorLanio, María Eliana
dc.creatorLomonte, Bruno
dc.creatorValle Garay, Aisel
dc.creatorÁlvarez Valcarcel, Carlos
dc.date.accessioned2020-01-22T20:20:00Z
dc.date.available2020-01-22T20:20:00Z
dc.date.issued2019
dc.identifier.citationhttps://www.sciencedirect.com/science/article/abs/pii/S0300908418302086?via%3Dihubes_ES
dc.identifier.issn0300-9084
dc.identifier.urihttp://hdl.handle.net/10669/80363
dc.description.abstractActinoporins constitute a unique class of pore-forming toxins found in sea anemones that being secreted as soluble monomers are able to bind and permeabilize membranes leading to cell death. The interest in these proteins has risen due to their high cytotoxicity that can be properly used to design immunotoxins against tumor cells and antigen-releasing systems to cell cytosol. In this work we describe a novel actinoporin produced by Anthopleura nigrescens, an anemone found in the Central American Pacific Ocean. Here we report the amino acid sequence of an actinoporin as deduced from cDNA obtained from total body RNA. The synthetic DNA sequence encoding for one cytolysin variant was expressed in BL21 Star (DE3) Escherichia coli and the protein purified by chromatography on CM Sephadex C-25 with more than 97% homogeneity as verified by MS-MS and HPLC analyses. This actinoporin comprises 179 amino acid residues, consistent with its observed isotope-averaged molecular mass of 19 661 Da. The toxin lacks Cys and readily permeabilizes erythrocytes, as well as L1210 cells. CD spectroscopy revealed that its secondary structure is dominated by beta structure (58.5%) with 5.5% of a-helix, and 35% of random structure. Moreover, binding experiments to lipidic monolayers and to liposomes, as well as permeabilization studies in vesicles, revealed that the affinity of this toxin for sphingomyelin-containing membranes is quite similar to sticholysin II (StII). Comparison by spectroscopic techniques and modeling the three-dimensional structure of nigrelysin (Ng) showed a high homology with StII but several differences were also detectable. Taken together, these results reinforce the notion that Ng is a novel member of the actinoporin pore-forming toxin (PFT) family with a HA as high as that of StII, the most potent actinoporin so far described, but with peculiar structural characteristics contributing to expand the understanding of the structure-function relationship in this protein family.es_ES
dc.description.sponsorshipInternational Foundation for Science/[F/5205–1]/IFS/Estados Unidoses_ES
dc.description.sponsorshipInternational Foundation for Science/[F/5205-2]/IFS/Estados Unidoses_ES
dc.description.sponsorshipUniversidad Nacional/[SIA 0067-13]/UNA/Costa Ricaes_ES
dc.description.sponsorshipUniversidad Nacional/[SIA 0116-16]/UNA/Costa Ricaes_ES
dc.language.isoen_USes_ES
dc.sourceBiochimie, vol.156, pp.206-223es_ES
dc.subjectActinoporines_ES
dc.subjectNigrelysines_ES
dc.subjectAnthopleura nigrescenses_ES
dc.subjectPore-forming toxines_ES
dc.subjectLytic activityes_ES
dc.subjectHemolysises_ES
dc.titleCloning, purification and characterization of nigrelysin, a novel actinoporin from the sea anemone Anthopleura nigrescenses_ES
dc.typeinfo:eu-repo/semantics/articlees_ES
dc.identifier.doi10.1016/j.biochi.2018.07.013
dc.description.procedenceUCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Instituto Clodomiro Picado (ICP)es_ES


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