Mostrar el registro sencillo del ítem

hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorTALBOT, Jean-Claude
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorDAUTANT, Alain
hal.structure.identifierLaboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels [LCBOSMV]
dc.contributor.authorPOLIDORI, Ange
hal.structure.identifierLaboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels [LCBOSMV]
dc.contributor.authorPUCCI, Bernard
hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorCOHEN-BOUACINA, Touria
hal.structure.identifierCentre de physique moléculaire optique et hertzienne [CPMOH]
dc.contributor.authorMAALI, A.
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorSALIN, Bénédicte
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorBRÈTHES, Daniel
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorVELOURS, Jean
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorGIRAUD, Marie-France
dc.date.issued2009
dc.identifier.issn0145-479X
dc.description.abstractEnLoss 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.isoen
dc.publisherSpringer Verlag
dc.subject.enMembrane protein complexes
dc.subject.enF1F0 ATP-synthase
dc.subject.enDetergent
dc.subject.enFluorinated surfactants
dc.title.enHydrogenated 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.typeArticle de revue
dc.identifier.doi10.1007/s10863-009-9235-5
dc.subject.halSciences du Vivant [q-bio]/Biologie cellulaire
bordeaux.journalJournal of Bioenergetics and Biomembranes
bordeaux.page349-360
bordeaux.volume41
bordeaux.issue4
bordeaux.peerReviewedoui
hal.identifierhal-00429653
hal.version1
hal.popularnon
hal.audienceInternationale
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-00429653v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Journal%20of%20Bioenergetics%20and%20Biomembranes&rft.date=2009&rft.volume=41&rft.issue=4&rft.spage=349-360&rft.epage=349-360&rft.eissn=0145-479X&rft.issn=0145-479X&rft.au=TALBOT,%20Jean-Claude&DAUTANT,%20Alain&POLIDORI,%20Ange&PUCCI,%20Bernard&COHEN-BOUACINA,%20Touria&rft.genre=article


Archivos en el ítem

ArchivosTamañoFormatoVer

No hay archivos asociados a este ítem.

Este ítem aparece en la(s) siguiente(s) colección(ones)

Mostrar el registro sencillo del ítem