Cyclohexyl-α maltoside as a highly efficient tool for membrane protein studies

Current Research in Structural Biology

Čeněk Kolar¹  Julie Winkel Missel²  Kamil Gotfryd²  Amalie Gerdt Laursen²  Nina Salustros²  Eva Ramos Becares²  Jonas Hyld Steffen²  Maria M. Garcia-Alai³  Angelica Struve Garcia³  Pontus Gourdon⁴ 

[1] Centre for Structural Systems Biology, Notkestrasse 85, D-22607, Hamburg, Germany;Department of Biomedical Sciences, Copenhagen University, Maersk Tower 7-9, Nørre Allé 14, DK-2200, Copenhagen N, Denmark;European Molecular Biology Laboratory Hamburg, Notkestrasse 85, D-22607, Hamburg, Germany;Glycon Biochemicals GmbH, Im Biotechnologie Park TGZ 1, D-14943, Luckenwalde, Germany;

关键词: Cryo-EM;    Crystallization;    Detergent;    Membrane proteins;    Solubilization;   

DOI  :  

来源: DOAJ

【 摘 要 】

Membrane proteins (MPs) constitute a large fraction of the proteome, but exhibit physicochemical characteristics that impose challenges for successful sample production crucial for subsequent biophysical studies. In particular, MPs have to be extracted from the membranes in a stable form. Reconstitution into detergent micelles represents the most common procedure in recovering MPs for subsequent analysis. n-dodecyl-β-D-maltoside (DDM) remains one of the most popular conventional detergents used in production of MPs. Here we characterize the novel DDM analogue 4-trans-(4-trans-propylcyclohexyl)-cyclohexyl α-maltoside (t-PCCαM), possessing a substantially lower critical micelle concentration (CMC) than the parental compound that represents an attractive feature when handling MPs. Using three different types of MPs of human and prokaryotic origin, i.e., a channel, a primary and a secondary active transporter, expressed in yeast and bacterial host systems, respectively, we investigate the performance of t-PCCαM in solubilization and affinity purification together with its capacity to preserve native fold and activity. Strikingly, t-PCCαM displays favorable behavior in extracting and stabilizing the three selected targets. Importantly, t-PCCαM promoted extraction of properly folded protein, enhanced thermostability and provided negatively-stained electron microscopy samples of promising quality. All-in-all, t-PCCαM emerges as competitive surfactant applicable to a broad portfolio of challenging MPs for downstream structure-function analysis.

【 授权许可】

Unknown