Please use this identifier to cite or link to this item: http://cmuir.cmu.ac.th/jspui/handle/6653943832/58348
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dc.contributor.authorPanchika Prangkioen_US
dc.contributor.authorSirikran Juntapremjiten_US
dc.contributor.authorMelanie Koehleren_US
dc.contributor.authorPeter Hinterdorferen_US
dc.contributor.authorChanan Angsuthanasombaten_US
dc.date.accessioned2018-09-05T04:23:00Z-
dc.date.available2018-09-05T04:23:00Z-
dc.date.issued2018-01-01en_US
dc.identifier.issn18755305en_US
dc.identifier.issn09298665en_US
dc.identifier.other2-s2.0-85045779856en_US
dc.identifier.other10.2174/0929866525666171201120456en_US
dc.identifier.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85045779856&origin=inwarden_US
dc.identifier.urihttp://cmuir.cmu.ac.th/jspui/handle/6653943832/58348-
dc.description.abstract© 2018 Bentham Science Publishers. Background: Adenylate cyclase (CyaA) is one of the major virulence factors of Bordetella pertussis that plays a key role in whooping cough pathogenesis. A putative transmembrane helical hairpin (α2-loop-α3), encompassing residues 529-594 of CyaA hemolysin (CyaA-Hly) domain, was previously proposed to be crucially involved in hemolytic activity against target erythrocytes. Objective: The main objective of this study was to gain more insight into membrane permeabilization of this toxin. Membrane-permeabilizing abilities of the purified 130-kDa CyaA-Hly and synthetic peptides corresponding to the helical component of interest, were evaluated. Methods: Synthetic peptides corresponding to the critical helical component, i.e. α2 (W528-G550), α3 (G568-R594) and α2-loop-α3 (W528-R594), were examined on various membrane models in comparison with the purified 130-kDa CyaA-Hly. The peptides were commercially synthesized and the purified toxin was obtained from recombinant plasmid construction and expression in Escherichia coli, followed by purification via immobilized-metal affinity chromatography. Membrane permeabilization or hemolysis of the peptides or the purified toxin were determined by liposomal leakage, hemolysis assays and atomic force microscopy (AFM) imaging. Results: Our results showed that the truncated 130-kDa CyaA-Hly, the synthetic peptides α2, α3 and the α2-loop-α3 hairpin exhibited distinct membrane-permeabilizing capacities in different membrane models. We demonstrated that the CyaA-Hly toxin and the peptides, especially the α2 peptide, caused nonspecific liposomal leakage as monitored by fluorescence dequenching of sulforhodamine B-loaded lipid vesicles. Notably, α2 peptide showed a predominant effect of membrane permeabilization when compared to α2-loop-α3 hairpin and α3 peptides. In addition, AFM imaging demonstrates lipid membrane disruption induced by the CyaA-Hly toxin or the peptidic α2-loop-α3 hairpin. Conclusion: Overall, the study provides the supporting evidence that the putative helical α2-loop-α3 hairpin could interact with the lipid membranes while the helical α2 peptide strongly induced liposomal leakage and hemolysis, as compared with the helical α3 or the α2-loop-α3 peptides, suggesting that the helix 2 from the hydrophobic region of CyaA-Hly is a crucial component that contributes to membrane permeabilization.en_US
dc.subjectBiochemistry, Genetics and Molecular Biologyen_US
dc.titleContributions of the hydrophobic helix 2 of the Bordetella pertussis CyaA-hemolysin to membrane permeabilizationen_US
dc.typeJournalen_US
article.title.sourcetitleProtein and Peptide Lettersen_US
article.volume25en_US
article.stream.affiliationsChiang Mai Universityen_US
article.stream.affiliationsBurapha Universityen_US
article.stream.affiliationsJohannes Kepler Universitat Linzen_US
article.stream.affiliationsUniversite Catholique de Louvainen_US
article.stream.affiliationsMahidol Universityen_US
article.stream.affiliationsBiophysics Institute for Research and Development (BIRD)en_US
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