Cyclohexyl-α maltoside as a highly efficient tool for membrane protein studies
(2021) In Current Research in Structural Biology 3. p.85-94- Abstract
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... (More)
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.
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- author
- Missel, Julie Winkel ; Salustros, Nina ; Becares, Eva Ramos ; Steffen, Jonas Hyld ; Laursen, Amalie Gerdt ; Garcia, Angelica Struve ; Garcia-Alai, Maria M. ; Kolar, Čeněk ; Gourdon, Pontus LU and Gotfryd, Kamil
- organization
- publishing date
- 2021
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Cryo-EM, Crystallization, Detergent, Membrane proteins, Solubilization
- in
- Current Research in Structural Biology
- volume
- 3
- pages
- 10 pages
- publisher
- Elsevier
- external identifiers
-
- scopus:85105688823
- pmid:34235488
- ISSN
- 2665-928X
- DOI
- 10.1016/j.crstbi.2021.03.002
- language
- English
- LU publication?
- yes
- id
- e7be1f09-fbaf-4bee-829b-5840b21c1a58
- date added to LUP
- 2021-06-07 15:27:18
- date last changed
- 2024-06-15 12:10:45
@article{e7be1f09-fbaf-4bee-829b-5840b21c1a58,
abstract = {{<p>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.</p>}},
author = {{Missel, Julie Winkel and Salustros, Nina and Becares, Eva Ramos and Steffen, Jonas Hyld and Laursen, Amalie Gerdt and Garcia, Angelica Struve and Garcia-Alai, Maria M. and Kolar, Čeněk and Gourdon, Pontus and Gotfryd, Kamil}},
issn = {{2665-928X}},
keywords = {{Cryo-EM; Crystallization; Detergent; Membrane proteins; Solubilization}},
language = {{eng}},
pages = {{85--94}},
publisher = {{Elsevier}},
series = {{Current Research in Structural Biology}},
title = {{Cyclohexyl-α maltoside as a highly efficient tool for membrane protein studies}},
url = {{http://dx.doi.org/10.1016/j.crstbi.2021.03.002}},
doi = {{10.1016/j.crstbi.2021.03.002}},
volume = {{3}},
year = {{2021}},
}