title: A class of mild surfactants that keep integral membrane proteins water-soluble for functional studies and crystallization.
abstract: Mixed protein-surfactant micelles are used for in vitro studies and 3D crystallization when solutions of pure, monodisperse integral membrane proteins are required. However, many membrane proteins undergo inactivation when transferred from the biomembrane into micelles of conventional surfactants with alkyl chains as hydrophobic moieties. Here we describe the development of surfactants with rigid, saturated or aromatic hydrocarbon groups as hydrophobic parts. Their stabilizing properties are demonstrated with three different integral membrane proteins. The temperature at which 50% of the binding sites for specific ligands are lost is used as a measure of stability and dodecyl-β-D-maltoside ('C12-b-M') as a reference for conventional surfactants. One surfactant increased the stability of two different G protein-coupled receptors and the human Patched protein receptor by approximately 10°C compared to C12-b-M. Another surfactant yielded the highest stabilization of the human Patched protein receptor compared to C12-b-M (13°C) but was inferior for the G protein-coupled receptors. In addition, one of the surfactants was successfully used to stabilize and crystallize the cytochrome b(6 )f complex from Chlamydomonas reinhardtii. The structure was solved to the same resolution as previously reported in C12-b-M.
authors.affiliation: Fachbereich Biologie, Universität Konstanz, Universitätsstr. 10, 78457 Konstanz, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Hovers
authors.forename: Meike
authors.initials: M
authors.lastname: Potschies
authors.forename: Ange
authors.initials: A
authors.lastname: Polidori
authors.forename: Bernard
authors.initials: B
authors.lastname: Pucci
authors.forename: Simon
authors.initials: S
authors.lastname: Raynal
authors.forename: Françoise
authors.initials: F
authors.lastname: Bonneté
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Serrano-Vega
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Tate
authors.forename: Daniel
authors.initials: D
authors.lastname: Picot
authors.forename: Yves
authors.initials: Y
authors.lastname: Pierre
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Popot
authors.forename: Rony
authors.initials: R
authors.lastname: Nehmé
authors.forename: Michel
authors.initials: M
authors.lastname: Bidet
authors.forename: Isabelle
authors.initials: I
authors.lastname: Mus-Veteau
authors.forename: Holger
authors.initials: H
authors.lastname: Busskamp
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Jung
authors.forename: Andreas
authors.initials: A
authors.lastname: Marx
authors.forename: Peter A
authors.initials: PA
authors.lastname: Timmins
authors.forename: Wolfram
authors.initials: W
authors.lastname: Welte
mesh_terms: D016825:Chlamydomonas reinhardtii / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D045346:Cytochrome b6f Complex / Q000737:chemistry; D005960:Glucosides / Q000737:chemistry; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D000072079:Patched Receptors; D011956:Receptors, Cell Surface / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry; D012995:Solubility; D013501:Surface-Active Agents / Q000737:chemistry*; D014867:Water / Q000737:chemistry*
chemical_list: D005960:Glucosides; D008565:Membrane Proteins; D000072079:Patched Receptors; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D013501:Surface-Active Agents; D014867:Water; C040358:dodecyl maltoside; D045346:Cytochrome b6f Complex