Asymmetric maltose neopentyl glycol amphiphiles for a membrane protein study: effect of detergent asymmetricity on protein stability

Bae, Hyoung Eun; Du, Yang; Hariharan, Parameswaran; Mortensen, Jonas S.; Kumar, Kaavya Krishna; Ha, Betty; Das, Manabendra; Lee, Hyun Sung; Løland, Claus J.; Guan, Lan; Kobilka, Brian K.; Chae, Pil Seok

doi:10.1039/c8sc02560f

Cited by 28 publications

(24 citation statements)

References 70 publications

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“…Hundreds more detergents have been developed to stabilise MPs further leading to HTP screens to identify the most useful [ 39 , 40 ]. Recently introduced detergents include the neopentyl glycols [ 41 ] and glyco-diosgenin (GDN) which along with digitonin account for around a third of single-particle cryo-EM (cryo-SPA) structures. The recently developed oligoglycerol class is useful for native mass spectrometry [ 42 ].…”

Section: Establishing a Purification Methodsmentioning

confidence: 99%

The high-throughput production of membrane proteins

Birch

Quigley

2021

Emerging Topics in Life Sciences

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Membrane proteins, found at the junctions between the outside world and the inner workings of the cell, play important roles in human disease and are used as biosensors. More than half of all therapeutics directly affect membrane protein function while nanopores enable DNA sequencing. The structural and functional characterisation of membrane proteins is therefore crucial. However, low levels of naturally abundant protein and the hydrophobic nature of membrane proteins makes production difficult. To maximise success, high-throughput strategies were developed that rely upon simple screens to identify successful constructs and rapidly exclude those unlikely to work. Parameters that affect production such as expression host, membrane protein origin, expression vector, fusion-tags, encapsulation reagent and solvent composition are screened in parallel. In this way, constructs with divergent requirements can be produced for a variety of structural applications. As structural techniques advance, sample requirements will change. Single-particle cryo-electron microscopy requires less protein than crystallography and as cryo-electron tomography and time-resolved serial crystallography are developed new sample production requirements will evolve. Here we discuss different methods used for the high-throughput production of membrane proteins for structural biology.

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Section: Establishing a Purification Methodsmentioning

confidence: 99%

The high-throughput production of membrane proteins

Birch

Quigley

2021

Emerging Topics in Life Sciences

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“…This analysis assumed that the binding and release cycle of Na + was in the fast-exchange regime. In addition, the CPMG dispersion profile was observed for 23 Na nuclear spins. Although the chemical shifts of Na + bound to A 2A AR and free Na + were similar (Figure 10b), their dispersion profiles indicated that they were not identical and that line broadening caused by intermediate exchange was taking place.…”

Section: ■ Monitoring Interactions Between Gpcrs and Their Environmentmentioning

confidence: 97%

Dynamic G Protein-Coupled Receptor Signaling Probed by Solution NMR Spectroscopy

Park

Lee

2020

Biochemistry

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Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool for investigating various dynamic features of G protein-coupled receptor (GPCR) signaling. In this Perspective, we focus on NMR techniques to characterize ligand-dependent conformational dynamics of GPCRs as well as the interaction of GPCRs with their environment and ligands. We also describe circumstances under which each technique should be applied, their advantages and disadvantages, and how they can be combined with other strategies to deepen the understanding of GPCR signaling at the molecular level.

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“…However, recently there has been a drive towards developing completely novel detergents that confer, in particular, elevated protein stability, compared with classical detergents. Examples of these include; a new class of steroid-based pentasaccharides [ 6 ], fluorinated glucose and maltose-based detergents [ 7 , 8 ], innovative facial amphiphiles (FAs) [ 9 , 10 ], 1,3,5-triazine-cored detergents [ 11 ], disulfide containing amphiphiles [ 12 ] and modified neopentyl glycol based detergents that have undergone subsequent modification to further improve their characteristics [ 13 ] ( Figure 2 ).…”

Section: Detergentsmentioning

confidence: 99%

Methods for the solubilisation of membrane proteins: the micelle-aneous world of membrane protein solubilisation

Ratkeviciute

Cooper

Knowles

2021

Biochemical Society Transactions

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The solubilisation of membrane proteins (MPs) necessitates the overlap of two contradictory events; the extraction of MPs from their native lipid membranes and their subsequent stabilisation in aqueous environments. Whilst the current myriad of membrane mimetic systems provide a range of modus operandi, there are no golden rules for selecting the optimal pipeline for solubilisation of a specific MP hence a miscellaneous approach must be employed balancing both solubilisation efficiency and protein stability. In recent years, numerous diverse lipid membrane mimetic systems have been developed, expanding the pool of available solubilisation strategies. This review provides an overview of recent developments in the membrane mimetic field, with particular emphasis placed upon detergents, polymer-based nanodiscs and amphipols, highlighting the latest reagents to enter the toolbox of MP research.

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Asymmetric maltose neopentyl glycol amphiphiles for a membrane protein study: effect of detergent asymmetricity on protein stability

Abstract: An asymmetric MNG, MNG-8,12, provided enhanced stability to human G protein-coupled receptors (GPCRs) compared to the symmetric MNG, MNG-3.

Cited by 28 publications

References 70 publications

The high-throughput production of membrane proteins

The high-throughput production of membrane proteins

Dynamic G Protein-Coupled Receptor Signaling Probed by Solution NMR Spectroscopy

Methods for the solubilisation of membrane proteins: the micelle-aneous world of membrane protein solubilisation

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