Cell-Free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-Synthase Subunits
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | LARRIEU, Isabelle | |
hal.structure.identifier | Centre de Recherche Paul Pascal [CRPP] | |
dc.contributor.author | TOLCHARD, James | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | SANCHEZ, Corinne | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | KONE, Edmond Yazo | |
hal.structure.identifier | Chimie et Biologie des Membranes et des Nanoobjets [CBMN] | |
dc.contributor.author | BARRAS, Alexandre | |
hal.structure.identifier | Centre de Recherche Paul Pascal [CRPP] | |
dc.contributor.author | STINES, Claire | |
hal.structure.identifier | Chimie et Biologie des Membranes et des Nanoobjets [CBMN] | |
dc.contributor.author | ODAERT, Benoît | |
hal.structure.identifier | Institut de biochimie et génétique cellulaires [IBGC] | |
dc.contributor.author | GIRAUD, Marie France | |
dc.date.accessioned | 2020-07-09T14:16:47Z | |
dc.date.available | 2020-07-09T14:16:47Z | |
dc.date.issued | 2017 | |
dc.identifier.uri | https://oskar-bordeaux.fr/handle/20.500.12278/10304 | |
dc.description.abstractEn | Small 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.en | Cell-Free Expression for the Study of Hydrophobic Proteins: The Example of Yeast ATP-Synthase Subunits | |
dc.type | Article de revue | |
dc.identifier.doi | 10.1007/978-1-4939-7151-0_4 | |
dc.subject.hal | Chimie/Matériaux | |
bordeaux.journal | Methods in molecular biology (Clifton, N.J.) | |
bordeaux.page | 57-90 | |
bordeaux.volume | 1635 | |
bordeaux.hal.laboratories | Institut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248 | * |
bordeaux.hal.laboratories | Institut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN, UMR 5248) | |
bordeaux.institution | Université de Bordeaux | |
bordeaux.institution | Bordeaux INP | |
bordeaux.COinS | ctx_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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