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Pore formation by equinqtoxin, a eukaryotic pore-forming toxin, requires a flexible N-terminal region and a stable beta-sandwichKristan, Katarina ...Actinoporins are eukaryotic pore-forming proteins that create 2-nm pores in natural and model lipid membranes by the self-association of four monomers. The regions that undergo conformational change ... and form part of the transmembrane pore are currently being defined. It was shown recently that theN-terminal region (residues 10-28) of equinatoxin, an actinoporin from Actinia equina, participates in building of the final pore wall. Assuming thatthe pore is formed solely by a polypeptide chain, other parts of the toxinshould constitute the conductive channel and here we searched for these regions by disulfide scanning mutagenesis. Only double cysteine mutants where the N-terminal segment 1-30 was attached to the (3-sandwich exhibited reduced hemolytic activity upon disulfide formation, showing that other parts of equinatoxin, particularly the beta-sandwich and importantly the C-terminal a-helix, do not undergo large conformational rearrangements during the pore formation.The role of the beta-sandwich stability was independently assessed via destabilization of a part of its hydrophobic core by mutations of the buried Trp117 These mutants were considerably less stable than the wild-type but exhibited similar or slightly lower permeabilizing activity. Collectively these results show that a flexible N-terminal region and stable beta-sandwich are pre-requisite for proper pore formation by the actinoporin family.Source: The Journal of biological chemistry. - ISSN 0021-9258 (Letn. 279, št. 45, 2004, str. 46509-46517)Type of material - article, component partPublish date - 2004Language - englishCOBISS.SI-ID - 18475481
Author
Kristan, Katarina |
Podlesek, Zdravko |
Hojnik, Vesna, biokemija |
Gutierrez-Aguirre, Ion |
Gunčar, Gregor |
Turk, Dušan, 1959 |
González-Mañas, M. |
Lakey, Jeremy H. |
Maček, Peter |
Anderluh, Gregor, 1969-
Topics
Sea Anemones |
Cnidarian Venoms |
Porins |
Mutation |
Binding Sites |
Cysteine |
Cloning, Molecular |
Hemolysins |
Disulfides |
Escherichia Coli |
Mesta vezave |
Morske vetrnice |
Mutacija |
Ožigalkarji, strupi |
Porini |
Cistein |
Disulfidi |
Escherichia coli |
Hemolizini |
Kloniranje molekulsko
source: The Journal of biological chemistry. - ISSN 0021-9258 (Letn. 279, št. 45, 2004, str. 46509-46517)
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Kristan, Katarina | 22470 |
Podlesek, Zdravko | 06902 |
Hojnik, Vesna, biokemija | |
Gutierrez-Aguirre, Ion | 27827 |
Gunčar, Gregor | 15639 |
Turk, Dušan, 1959 | 04988 |
González-Mañas, M. | |
Lakey, Jeremy H. | |
Maček, Peter | 06994 |
Anderluh, Gregor, 1969- | 15686 |
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