Shiga toxin B-subunit (STxB), a protein involved in the cell-binding and intracellular trafficking of Shiga holotoxin, binds to a specific glycolipid, the globotriaosyl ceramide (Gb(3)) Tryptophan residues of STxB, located at the protein-membrane interface, allow one to study its interaction with model membranes by means of spectroscopic methods with no need for chemical derivatisation with a fluorophore. The protein emits maximally around 346 nm and a blue shift of about 8 mn, as well as the occurrence of changes in the emission fluorescence intensity spectra, is indicative of insertion and partition into the membrane. However, the interaction seems to take place without pentamer dissociation. Acrylamide quenching experiments confirm tryptophan residues become less exposed to solvent when in the presence of vesicles, and the use of lipophilic probes suggests that they are located in a shallow position near the water/membrane interface. Fluorescence intensity and lifetime measurements upon STxB titration with Gb(3)-containing vesicles suggest a complex STxB/Gb(3) docking mechanism involving static quenching in the later stages. Based on our observations, a model of the protein-membrane interaction is proposed and the STxB membrane partition and binding constants were calculated. (c) 2007 Elsevier B.V. All rights reserved.
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Garcia-Castillo, Maria Daniela
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Thi Tran
Bobard, Alexandre
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Inst Pasteur, Dynam Interact Hote Pathogene, F-75724 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Bobard, Alexandre
Renard, Henri-Francois
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Renard, Henri-Francois
Rathjen, Stefan J.
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Rathjen, Stefan J.
Dransart, Estelle
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Dransart, Estelle
Stechmann, Bahne
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Stechmann, Bahne
Lamaze, Christophe
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CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, France
Inst Curie, Ctr Rech, Membrane Dynam & Mech Intracellular Signaling Grp, F-75248 Paris 05, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Lamaze, Christophe
Lord, Mike
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Univ Warwick, Dept Biol Sci, Coventry CV4 7AL, W Midlands, EnglandPSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Lord, Mike
Cintrat, Jean-Christophe
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CEA Saclay, CEA, LabEx LERMIT, IBiTec S SCBM, F-91191 Gif Sur Yvette, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Cintrat, Jean-Christophe
Enninga, Jost
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Inst Pasteur, Dynam Interact Hote Pathogene, F-75724 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
Enninga, Jost
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Tartour, Eric
Johannes, Ludger
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PSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France
CNRS, UMR3666, F-75005 Paris, France
INSERM, U1143, F-75005 Paris, FrancePSL Res Univ, Inst Curie, Endocyt Trafficking & Therapeut Delivery Grp, F-75248 Paris 05, France