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Two-step membrane binding by equinatoxin II, a pore-forming toxin from the seaanemone, involves and exposed aromatic cluster and a flexible helixHong, Qi ...Equinatoxin II (EqtII) belongs to a unique family of 20-kDa pore-forming toxins from sea anemones. These toxins preferentially bind to membranes containing sphingomyelin and create cation-selective ... pores by oligomerization of 3-4 monomers. In this work we have studied the binding of EqtII to lipid membranes by the use of lipid monolayers and surface plasmon resonance (SPR). The binding is a two-step process, separately mediated by two regions of the molecule. An exposed aromatic cluster involving tryptophans 112 and 116 mediates the initial attachment that is prerequisite for the next step. Stericshielding of the aromatic cluster or mutation of Trp-112 and -116 to phenylalanine significantly reduces the toxin-lipid interaction. The second step is promoted by the N-terminal amphiphilic helix, which translocates into the lipid phase. The two steps were distinguished by the use of a double cysteine mutant having the N-terminal helix fixed to the protein core by a disulfide bond. The kinetics of membrane binding derived from the SPR experiments could be fitted to a two-stage binding model. Finally, by using membraneembedded quenchers, we showed that EqtII does not insert deeply in themembrane. The first step of the EqtII binding is reminiscent of the bindingof the evolutionarily distant cholesterol-dependant cytolysins, which share a similar structural motif in the membrane attachment domain.Vir: The Journal of biological chemistry. - ISSN 0021-9258 (Letn. 277, Št. 44, 2002, str. 41916-41924)Vrsta gradiva - članek, sestavni delLeto - 2002Jezik - angleškiCOBISS.SI-ID - 15480793
Avtor
Hong, Qi |
Gutiérrez-Aguirres, Ion |
Barlič, Ariana |
Malovrh, Petra |
Kristan, Katarina |
Podlesek, Zdravko |
Maček, Peter |
Turk, Dušan |
Gonzales-Manas, Juan M. |
Lakey, Jeremy H. |
Anderluh, Gregor, 1969-
Teme
Cnidarian Venoms |
Cytotoxins |
Porins |
Lipid Bilayers |
Binding Sites |
Sea Anemones |
Avidin |
Hemolysins |
Tryptophan |
Cysteine |
Citotoksini |
Lipidna, dvojna plast |
Mesta vezave |
Ožigalkarji, strupi |
Porini |
Avidin |
Cistein |
Hemolizini |
Morske vetrnice |
Triptofan
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vir: The Journal of biological chemistry. - ISSN 0021-9258 (Letn. 277, Št. 44, 2002, str. 41916-41924)
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Povezave do osebnih bibliografij avtorjev | Povezave do podatkov o raziskovalcih v sistemu SICRIS |
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Barlič, Ariana | 15627 |
Malovrh, Petra | 18203 |
Kristan, Katarina | 22470 |
Podlesek, Zdravko | 06902 |
Maček, Peter | 06994 |
Turk, Dušan | 04988 |
Anderluh, Gregor, 1969- | 15686 |
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