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Cell-Free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-Synthase Subunits

hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorLARRIEU, Isabelle
hal.structure.identifierCentre de Recherche Paul Pascal [CRPP]
dc.contributor.authorTOLCHARD, James
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorSANCHEZ, Corinne
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorKONE, Edmond Yazo
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorBARRAS, Alexandre
hal.structure.identifierCentre de Recherche Paul Pascal [CRPP]
dc.contributor.authorSTINES, Claire
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorODAERT, Benoît
hal.structure.identifierInstitut de biochimie et génétique cellulaires [IBGC]
dc.contributor.authorGIRAUD, Marie France
dc.date.accessioned2020-07-09T14:16:47Z
dc.date.available2020-07-09T14:16:47Z
dc.date.issued2017
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/10304
dc.description.abstractEnSmall hydrophobic membrane proteins or proteins with hydrophobic domains are often difficult to produce in bacteria. The cell-free expression system was found to be a very good alternative for the expression of small hydrophobic subunits of the yeast ATP-synthase, such as subunits e, g, k, i, f and the membrane domain of subunit 4, proteins that are suspected to play a role in the stability of ATP-synthase dimers. All of these proteins could be produced in milligrams amounts using the cell-free "precipitate mode" and were successfully solubilized in the presence of lysolipid 1-myristoyl-2-hydroxy-sn-glycero-3-phospho-1'-rac-glycerol. Purified proteins were also found suitable for structural investigations. An example is given with the NMR backbone assignment of the isotopically labeled subunit g. Protocols are also described for raising specific polyclonal antibodies against overexpressed cell-free proteins.
dc.title.enCell-Free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-Synthase Subunits
dc.typeArticle de revue
dc.identifier.doi10.1007/978-1-4939-7151-0_4
dc.subject.halChimie/Matériaux
bordeaux.journalMethods in molecular biology (Clifton, N.J.)
bordeaux.page57-90
bordeaux.volume1635
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248*
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN, UMR 5248)
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionBordeaux INP
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=Methods%20in%20molecular%20biology%20(Clifton,%20N.J.)&rft.date=2017&rft.volume=1635&rft.spage=57-90&rft.epage=57-90&rft.au=LARRIEU,%20Isabelle&TOLCHARD,%20James&SANCHEZ,%20Corinne&KONE,%20Edmond%20Yazo&BARRAS,%20Alexandre&rft.genre=article


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