Releasing the technical ‘shackles’ on GPCR drug discovery: opportunities enabled by detergent-free polymer lipid particle (PoLiPa) purification

Hothersall, Joanne; Jones, Andrew Y.; Dafforn, Timothy R.; Perrior, Trevor; Chapman, Kathryn

doi:10.1016/j.drudis.2020.08.006

Cited by 10 publications

(6 citation statements)

References 93 publications

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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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“…Recent studies have demonstrated the nanodisc formation by a variety of polymers that are cationic, − anionic, ,− zwitterionic, styrene-containing, styrene-free, ,, charge-free, and metal-chelators . Some of these polymers have been shown to be stable against pH, and some of them can be used to form nanodiscs for certain pH values. ,,, This library of amphipathic polymers has significantly expanded the applications of nanodiscs. − …”

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confidence: 99%

Solid-State NMR Study to Probe the Effects of Divalent Metal Ions (Ca²⁺ and Mg²⁺) on the Magnetic Alignment of Polymer-Based Lipid Nanodiscs

2021

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Divalent cations, especially Ca2+ and Mg2+, play a vital role in the function of biomolecules and making them important to be constituents in samples for in vitro biophysical and biochemical characterizations. Although lipid nanodiscs are becoming valuable tools for structural biology studies on membrane proteins and for drug delivery, most types of nanodiscs used in these studies are unstable in the presence of divalent metal ions. To avoid the interaction of divalent metal ions with the belt of the nanodiscs, synthetic polymers have been designed and demonstrated to form stable lipid nanodiscs under such unstable conditions. Such polymer-based nanodiscs have been shown to provide an ideal platform for structural studies using both solid-state and solution NMR spectroscopies because of the near-native cell-membrane environment they provide and the unique magnetic-alignment behavior of large-size nanodiscs. In this study, we report an investigation probing the effects of Ca2+ and Mg2+ ions on the formation of polymer-based lipid nanodiscs and the magnetic-alignment properties using a synthetic polymer, styrene maleimide quaternary ammonium (SMA-QA), and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) lipids. Phosphorus-31 NMR experiments were used to evaluate the stability of the magnetic-alignment behavior of the nanodiscs for varying concentrations of Ca2+ or Mg2+ at different temperatures. It is remarkable that the interaction of divalent cations with lipid headgroups promotes the stacking up of nanodiscs that results in the enhanced magnetic alignment of nanodiscs. Interestingly, the reported results show that both the temperature and the concentration of divalent metal ions can be optimized to achieve the optimal alignment of nanodiscs in the presence of an applied magnetic field. We expect the reported results to be useful in the design of nanodisc-based nanoparticles for various applications in addition to atomic-resolution structural and dynamics studies using NMR and other biophysical techniques.

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“…[7] While progress has been made in enhancing expression and structural stability of GPCRs, [8] the accessibility of target material for discovery programs, both using small molecule or antibody approaches, remains a major bottleneck. [9] of those of protein-protein interactions, which have long been considered difficult to modulate using small molecules. [13,14] At the same time, they are too small to be fully engaged using antibodies, which generally interact with the extracellular domain of the receptor only.…”

Section: Obtaining Purified Gpcrs For Screeningmentioning

confidence: 99%

Precision-engineered Peptide and Protein Analogs: Establishing a New Discovery Platform for Potent GPCR Modulators

Akondi

Paolini-Bertrand

Hartley

2021

Chimia

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Numerous members of the human G protein-coupled receptor (GPCR) superfamily are receptors of therapeutic interest. GPCRs are considered to be highly tractable for drug discovery, representing the targets of approximately one-third of currently licensed drugs. These successful drug discovery outcomes cover only a relatively small subset of the superfamily, however, and many other attractive receptors have proven to present significant challenges. Among these difficult GPCRs are those whose natural ligands are peptides and proteins. In this review we explain the obstacles faced by GPCR drug discovery campaigns, with particular focus on those related to peptide and protein GPCRs. We describe a novel and promising approach for these targets based on engineering of their natural ligands and describe an integrated discovery platform that allows potent ligand analogs to be discovered rapidly and efficiently. Finally, we present a case study involving the chemokine receptor CCR5 to show that this approach can be used to generate new drugs for peptide and protein GPCR targets combining best-in-class potency with tunable signaling activity.

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Releasing the technical ‘shackles’ on GPCR drug discovery: opportunities enabled by detergent-free polymer lipid particle (PoLiPa) purification

Cited by 10 publications

References 93 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

Solid-State NMR Study to Probe the Effects of Divalent Metal Ions (Ca²⁺ and Mg²⁺) on the Magnetic Alignment of Polymer-Based Lipid Nanodiscs

Precision-engineered Peptide and Protein Analogs: Establishing a New Discovery Platform for Potent GPCR Modulators

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Releasing the technical ‘shackles’ on GPCR drug discovery: opportunities enabled by detergent-free polymer lipid particle (PoLiPa) purification

Cited by 10 publications

References 93 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

Solid-State NMR Study to Probe the Effects of Divalent Metal Ions (Ca2+ and Mg2+) on the Magnetic Alignment of Polymer-Based Lipid Nanodiscs

Precision-engineered Peptide and Protein Analogs: Establishing a New Discovery Platform for Potent GPCR Modulators

Contact Info

Product

Resources

About

Solid-State NMR Study to Probe the Effects of Divalent Metal Ions (Ca²⁺ and Mg²⁺) on the Magnetic Alignment of Polymer-Based Lipid Nanodiscs