Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid

Harwood, Clare R.; Sykes, David A.; Hoare, Bradley L.; Heydenreich, Franziska M.; Uddin, Romez; Poyner, David R.; Briddon, Stephen J.; Veprintsev, Dmitry B.

doi:10.1016/j.isci.2021.103362

Cited by 11 publications

(8 citation statements)

References 36 publications

(43 reference statements)

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“…SMA and related copolymers found many diverse applications in structural biology. First of all, they were efficient in solubilization of a wide range of membrane proteins, including GPCRs [ 107 , 108 , 109 , 110 , 111 , 112 ], ABC transporters [ 113 , 114 , 115 , 116 ], ion channels [ 117 , 118 , 119 ], photoreaction centers [ 61 , 120 , 121 ], and electron transport chain complexes [ 62 ], expressed in bacteria [ 42 , 122 , 123 , 124 , 125 ], yeast [ 126 , 127 , 128 , 129 ], insect [ 116 , 130 , 131 ], and mammalian cells [ 115 , 132 , 133 ], as well as plants [ 134 ]. As reviewed by Overduin and Esmaili, SMA is effective for solubilizing both monomeric and oligomeric proteins as well as those that are unstable, low-abundance, or lipid-dependent [ 13 ].…”

Section: Applications Of Lipodiscs In Structural Biologymentioning

confidence: 99%

Mechanisms of Formation, Structure, and Dynamics of Lipoprotein Discs Stabilized by Amphiphilic Copolymers: A Comprehensive Review

et al. 2022

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Amphiphilic copolymers consisting of alternating hydrophilic and hydrophobic units account for a major recent methodical breakthrough in the investigations of membrane proteins. Styrene–maleic acid (SMA), diisobutylene–maleic acid (DIBMA), and related copolymers have been shown to extract membrane proteins directly from lipid membranes without the need for classical detergents. Within the particular experimental setup, they form disc-shaped nanoparticles with a narrow size distribution, which serve as a suitable platform for diverse kinds of spectroscopy and other biophysical techniques that require relatively small, homogeneous, water-soluble particles of separate membrane proteins in their native lipid environment. In recent years, copolymer-encased nanolipoparticles have been proven as suitable protein carriers for various structural biology applications, including cryo-electron microscopy (cryo-EM), small-angle scattering, and conventional and single-molecule X-ray diffraction experiments. Here, we review the current understanding of how such nanolipoparticles are formed and organized at the molecular level with an emphasis on their chemical diversity and factors affecting their size and solubilization efficiency.

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Section: Applications Of Lipodiscs In Structural Biologymentioning

confidence: 99%

Mechanisms of Formation, Structure, and Dynamics of Lipoprotein Discs Stabilized by Amphiphilic Copolymers: A Comprehensive Review

et al. 2022

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“…Thus, it substantially goes beyond previous work in which nanodiscs from cell extracts were reconstituted after initial detergent-mediated solubilization of prokaryotic membrane proteins. 44 Finally, with recent advances in polymer technology to isolate and characterize membrane proteins in their functional states, 45,46 this work can serve as a platform for future experiments with polymer-encapsulated nanodiscs of superior properties or more specific polymers that target specific cellular compartments.…”

Section: ■ Conclusionmentioning

confidence: 99%

Cryo-Electron Microscopy Snapshots of Eukaryotic Membrane Proteins in Native Lipid-Bilayer Nanodiscs

et al. 2022

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New technologies for purifying membrane-bound protein complexes in combination with cryo-electron microscopy (EM) have recently allowed the exploration of such complexes under near-native conditions. In particular, polymer-encapsulated nanodiscs enable the study of membrane proteins at high resolution while retaining protein−protein and protein−lipid interactions within a lipid bilayer. However, this powerful technology has not been exploited to address the important question of how endogenous�as opposed to overexpressed�membrane proteins are organized within a lipid environment. In this work, we demonstrate that biochemical enrichment protocols for native membrane−protein complexes from Chaetomium thermophilum in combination with polymer-based lipid-bilayer nanodiscs provide a substantial improvement in the quality of recovered endogenous membrane−protein complexes. Mass spectrometry results revealed ∼1123 proteins, while multiple 2D class averages and two 3D reconstructions from cryo-EM data furnished prominent structural signatures. This integrated methodological approach to enriching endogenous membrane−protein complexes provides unprecedented opportunities for a deeper understanding of eukaryotic membrane proteomes.

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“…DIBMA has also been successfully utilised to extract a GPCR, the β 2 ‐adrenergic receptor (β 2 AR), from membranes, albeit with a solubilisation efficiency about a third of that achieved with DDM. The extracted protein had similar ligand‐binding characteristics in both DIBMA and DDM but was more stable in DIBMA [64] . Electroneutral sulfo‐DIBMA has been used to explore lipid exchange between polymer environments, and this agent appears to be less susceptible to changes in local ion concentrations than DIBMA [65] …”

Section: Polymersmentioning

confidence: 99%

“…The extracted protein had similar ligand-binding characteristics in both DIBMA and DDM but was more stable in DIBMA. [64] Electroneutral sulfo-DIBMA has been used to explore lipid exchange between polymer environments, and this agent appears to be less susceptible to changes in local ion concentrations than DIBMA. [65] Inulins are an additional group of nanodisc forming polymers to have been recently reported.…”

Section: Polymersmentioning

confidence: 99%

Advances in Membrane Mimetic Systems for Manipulation and Analysis of Membrane Proteins: Detergents, Polymers, Lipids and Scaffolds

Woubshete,

Cioccolo,

Byrne

2024

ChemPlusChem

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Extracting membrane proteins from the hydrophobic environment of the biological membrane, in a physiologically relevant and stable state, suitable for downstream analysis remains a challenge. The traditional route to membrane protein extraction has been to use detergents and the last 15 years or so have seen a veritable explosion in the development of novel detergents with improved properties, making them more suitable for individual proteins and specific applications. There have also been significant advances in the development of encapsulation of membrane proteins in lipid based nanodiscs, either directly from the native membrane using polymers allowing effective capture of the protein and protein‐associated membrane lipids, or via reconstitution of detergent extracted and purified protein into nanodiscs of defined lipid composition. All of these advances have been successfully applied to the study of membrane proteins via a range of techniques and there have been some spectacular membrane protein structures solved using these new approaches. In addition, the first detailed structural and biophysical analyses of membrane proteins retained within a biological membrane have been reported. Here we summarise and review the recent advances with respect to these new agents and systems for membrane protein extraction, reconstitution and analysis.

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Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid

Cited by 11 publications

References 36 publications

Mechanisms of Formation, Structure, and Dynamics of Lipoprotein Discs Stabilized by Amphiphilic Copolymers: A Comprehensive Review

Mechanisms of Formation, Structure, and Dynamics of Lipoprotein Discs Stabilized by Amphiphilic Copolymers: A Comprehensive Review

Cryo-Electron Microscopy Snapshots of Eukaryotic Membrane Proteins in Native Lipid-Bilayer Nanodiscs

Advances in Membrane Mimetic Systems for Manipulation and Analysis of Membrane Proteins: Detergents, Polymers, Lipids and Scaffolds

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