Evaluation of different approaches used to study membrane permeabilization by actinoporins on model lipid vesicles

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dc.contributor.authorPalacios Ortega, Juan
dc.contributor.authorRivera-de-Torre, Esperanza
dc.contributor.authorGavilanes, José G.
dc.contributor.authorSlotte, J. Peter
dc.contributor.authorMartínez Del Pozo, Álvaro
dc.date.accessioned2023-06-16T15:20:05Z
dc.date.available2023-06-16T15:20:05Z
dc.date.issued2020-04-21
dc.description.abstractRelease of aqueous contents from model lipid vesicles has been a standard procedure to evaluate pore formation efficiency by actinoporins, such as sticholysin II (StnII), for the last few decades. However, regardless of the probe of choice, the results reported that StnII action was never able to empty the vesicles completely. This was hard to explain if StnII pores were to be stable and always leaky for the probes used. To address this question, we have used a variety of probes, including rhodamine 6G or Tb3+, to test the permeability of StnII's pores. Our results indicate that calcein was in fact too large to fit through StnII's pores, and that the standard method in the field is actually reporting StnII-induced transient permeation of the membrane rather than the passage of solutes through the stable assembled pores. In order to evaluate the permeability of these structures, we used a dithionite-based assay, which showed that the final pores were in fact open. Thus, our results indicate that the stable actinoporins' pores are open in spite of plateaued classic release curves. Besides the proper pore, the first stages of pore formation would inflict serious damage to living cells as well.
dc.description.departmentSección Deptal. de Bioquímica y Biología Molecular (Biológicas)
dc.description.facultyFac. de Ciencias Biológicas
dc.description.refereedTRUE
dc.description.sponsorshipUniversidad Complutense de Madrid (UCM)/Banco de Santander
dc.description.sponsorshipISB/ÅA Åbo Akademi University, Turku, Finland
dc.description.sponsorshipSigrid Jusélius Foundation
dc.description.sponsorshipJane and Aatos Erkko Foundation
dc.description.sponsorshipMagnus Ehrnrooth Foundation
dc.description.statuspub
dc.eprint.idhttps://eprints.ucm.es/id/eprint/61441
dc.identifier.doi10.1016/j.bbamem.2020.183311
dc.identifier.issn0005-2736
dc.identifier.officialurlhttps://www.sciencedirect.com/science/article/abs/pii/S0005273620301425
dc.identifier.urihttps://hdl.handle.net/20.500.14352/6388
dc.issue.number9
dc.journal.titleBiochimica et Biophysica Acta (BBA) - Biomembranes
dc.language.isoeng
dc.page.final8
dc.page.initial1
dc.publisherElsevier
dc.relation.projectIDPR75/18- 21561
dc.rights.accessRightsrestricted access
dc.subject.cdu577.112
dc.subject.keywordPore-forming-proteins
dc.subject.keywordLeakage Calcein
dc.subject.keywordRhodamine
dc.subject.keywordTerbium
dc.subject.keywordNBD-dithionite
dc.subject.ucmBioquímica (Biología)
dc.subject.unesco2302 Bioquímica
dc.titleEvaluation of different approaches used to study membrane permeabilization by actinoporins on model lipid vesicles
dc.typejournal article
dc.volume.number1862
dspace.entity.typePublication
relation.isAuthorOfPublication4d35a8a6-8bd3-4ff4-b179-57581d8d36d8
relation.isAuthorOfPublication.latestForDiscovery4d35a8a6-8bd3-4ff4-b179-57581d8d36d8
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