Hydrogenated and fluorinated surfactants derived from Tris (hydroxymethyl)-acrylamidomethane allow the purification of a highly active yeast F1-F0 ATP-synthase with an enhanced stability
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | TALBOT, Jean-Claude | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | DAUTANT, Alain | |
hal.structure.identifier | Laboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels [LCBOSMV] | |
dc.contributor.author | POLIDORI, Ange | |
hal.structure.identifier | Laboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels [LCBOSMV] | |
dc.contributor.author | PUCCI, Bernard | |
hal.structure.identifier | Centre de physique moléculaire optique et hertzienne [CPMOH] | |
dc.contributor.author | COHEN-BOUACINA, Touria | |
hal.structure.identifier | Centre de physique moléculaire optique et hertzienne [CPMOH] | |
dc.contributor.author | MAALI, A. | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | SALIN, Bénédicte | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | BRÈTHES, Daniel | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | VELOURS, Jean | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | GIRAUD, Marie-France | |
dc.date.issued | 2009 | |
dc.identifier.issn | 0145-479X | |
dc.description.abstractEn | Loss of stability and integrity of large membrane protein complexes as well as their aggregation in a non-lipidic environment are the major bottlenecks to their structural studies. We have tested C12H25-S-poly-Tris-(hydroxymethyl)acrylamidomethane (H12-TAC) among many other detergents for extracting the yeast F1F0 ATP-synthase. H12-TAC was found to be a very efficient detergent for removing the enzyme from mitochondrial membranes without altering its sensitivity towards specific ATP-synthase inhibitors. This extracted enzyme was then solubilized by either dodecyl maltoside (DDM), H12-TAC or fluorinated surfactants such as C2H5-C6F12-C2H4-S-poly-Tris-(hydroxymethyl)acrylamidomethane (H2F6-TAC) or C6F13-C2H4-S-poly-Tris-(hydroxymethyl)acrylamidomethane (F6-TAC), two surfactants exhibiting a comparable polar head to H12-TAC but bearing a fluorinated hydrophobic tail. Preparations from enzymes purified in the presence of H12-TAC were found to be more adapted for AFM imaging than ATP-synthase purified with DDM. Keeping H12-TAC during the Ni-NTA IMAC purification step or replacing it by DDM at low concentrations did not however allow preserving enzyme activity, while fluorinated surfactants H2F6-TAC and F6-TAC were found to enhance enzyme stability and integrity as indicated by sensitivity towards inhibitors. ATPase specific activity was higher with F6-TAC than with H2F6-TAC. When enzymes were mixed with egg phosphatidylcholine, ATP-synthases purified in the presence of H2F6-TAC or F6-TAC were more stable upon time than the DDM purified enzyme. Furthermore, in the presence of lipids, an activation of ATP-synthases was observed that was transitory for enzymes purified with DDM, but lasted for weeks for ATP-synthases isolated in the presence of molecules with Tris polyalcoholic moieties. Relipidated enzymes prepared with fluorinated surfactants remained highly sensitive towards inhibitors, even after 6 weeks. | |
dc.language.iso | en | |
dc.publisher | Springer Verlag | |
dc.subject.en | Membrane protein complexes | |
dc.subject.en | F1F0 ATP-synthase | |
dc.subject.en | Detergent | |
dc.subject.en | Fluorinated surfactants | |
dc.title.en | Hydrogenated and fluorinated surfactants derived from Tris (hydroxymethyl)-acrylamidomethane allow the purification of a highly active yeast F1-F0 ATP-synthase with an enhanced stability | |
dc.type | Article de revue | |
dc.identifier.doi | 10.1007/s10863-009-9235-5 | |
dc.subject.hal | Sciences du Vivant [q-bio]/Biologie cellulaire | |
bordeaux.journal | Journal of Bioenergetics and Biomembranes | |
bordeaux.page | 349-360 | |
bordeaux.volume | 41 | |
bordeaux.issue | 4 | |
bordeaux.peerReviewed | oui | |
hal.identifier | hal-00429653 | |
hal.version | 1 | |
hal.popular | non | |
hal.audience | Internationale | |
hal.origin.link | https://hal.archives-ouvertes.fr//hal-00429653v1 | |
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