Vibrio cholerae Hemolysin Implication of Amphiphilicity and Lipid-Induced Conformational Change for its Pore-forming Activity
EPrints@IICB
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Title |
Vibrio cholerae Hemolysin Implication of Amphiphilicity and Lipid-Induced Conformational Change for its Pore-forming Activity |
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Creator |
Chattopadhyay, Kausik
Bhattacharyya, Debasish Banerjee, Kalyan K |
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Subject |
Structural Biology & Bioinformatics
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Description |
Vibrio cholerae hemolysin (HlyA), a water-soluble protein with a native monomeric relative molecular mass of 65 000, forms transmembrane pentameric channels in target biomembranes. T heHlyA binds to lipid vesicles nonspecifically and without saturation; however, self-assembly is triggered specifically by cholesterol.Here we show that the HlyA partitioned quantitatively to amphiphilic media irrespective of their compositions, indicating that the toxin had an amphiphilic surface.Asialofe tuin, a b1-galactosyl-terminated glycoprotein, which binds specifically to the HlyA in a lectin-glycoprotein type of interaction and inhibits carbohydrate- independent interaction of the toxin with lipid, reduced effective amphiphilicity of the toxin significantly. Resistance of the HlyA to proteases together with the tryptophan fluorescence emission spectrum suggested a compact structure for the toxin.Fluorescence energy transfer from the HlyA to dansyl-phosphatidylethanolamine required the presence of cholesterol in the lipid bilayer and was synchronous with oligomerization.Phospholipid bilayer without cholesterol caused a partial unfolding of the HlyA monomer as indicated by the transfer of tryptophan residues from the nonpolar core of the protein to amore polar region. These observations suggested: (a) partitioning of the HlyA to lipid vesicles is driven by the tendency of the amphiphilic toxin to reduce energetically unfavorable contacts with water and is not affected significantly by the composition of the vesicles; and (b) partial unfolding of the HlyA at the lipid– water interface precedes and promotes cholesterol-induced oligomerization to an insertion-competent configuration. |
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Publisher |
Blackwell Publishing
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Date |
2002
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Type |
Article
PeerReviewed |
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Format |
application/pdf
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Identifier |
http://www.eprints.iicb.res.in/754/1/EUROPEAN_JOURNAL_OF_BIOCHEMISTRY_269(_17)_4351%2D4358;2002[29].pdf
Chattopadhyay, Kausik and Bhattacharyya, Debasish and Banerjee, Kalyan K (2002) Vibrio cholerae Hemolysin Implication of Amphiphilicity and Lipid-Induced Conformational Change for its Pore-forming Activity. European Journal Of Biochemistry, 269 (17). pp. 4351-4358. |
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Relation |
http://dx.doi.org/10.1046/j.1432-1033.2002.03137.x
http://www.eprints.iicb.res.in/754/ |
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