Nanodiscs for structural and functional studies of membrane proteins

Denisov, Ilia G.; Sligar, Stephen G.

doi:10.1038/nsmb.3195

Cited by 432 publications

(397 citation statements)

References 72 publications

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“…Methods for reconstituting membrane proteins in lipid-based nanoparticles (liposomes) or high-density apolipoprotein particles (nanodiscs) have been offered as a possible solution for these issues [46,48,49]. Such approaches have been used routinely for biochemical, biophysical, and structural studies of membrane proteins.…”

Section: Advantages Of Evs Over Other Nanoparticles From the View Of mentioning

confidence: 99%

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Yang

Hong

Cho

et al. 2018

J of Extracellular Vesicle

187

148

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Membrane proteins are of great research interest, particularly because they are rich in targets for therapeutic application. The suitability of various membrane proteins as targets for therapeutic formulations, such as drugs or antibodies, has been studied in preclinical and clinical studies. For therapeutic application, however, a protein must be expressed and purified in as close to its native conformation as possible. This has proven difficult for membrane proteins, as their native conformation requires the association with an appropriate cellular membrane. One solution to this problem is to use extracellular vesicles as a display platform. Exosomes and microvesicles are membranous extracellular vesicles that are released from most cells. Their membranes may provide a favourable microenvironment for membrane proteins to take on their proper conformation, activity, and membrane distribution; moreover, membrane proteins can cluster into microdomains on the surface of extracellular vesicles following their biogenesis. In this review, we survey the state-of-the-art of extracellular vesicle (exosome and small-sized microvesicle)-based therapeutics, evaluate the current biological understanding of these formulations, and forecast the technical advances that will be needed to continue driving the development of membrane protein therapeutics.

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Section: Advantages Of Evs Over Other Nanoparticles From the View Of mentioning

confidence: 99%

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Yang

Hong

Cho

et al. 2018

J of Extracellular Vesicle

187

148

View full text Add to dashboard Cite

show abstract

“…In contrast, lipid nanodiscs, which were introduced by Sligar and co‐workers,2 appear to be a promising solution for the aforementioned drawbacks of micelles and liposomes 9. Nanodisc technology12 has emerged as a new technique to purify and reconstitute proteins in a detergent‐free lipid‐bilayer environment 11, 36. A comparison of membrane protein dynamics in micelles and nanodiscs revealed that micelles compromised the inherent flexibility of the protein, despite conservation of the structure 21, 28.…”

Section: Introductionmentioning

confidence: 99%

Lipid Internal Dynamics Probed in Nanodiscs

et al. 2017

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Nanodiscs offer a very promising tool to incorporate membrane proteins into native‐like lipid bilayers and an alternative to liposomes to maintain protein functions and protein–lipid interactions in a soluble nanoscale object. The activity of the incorporated membrane protein appears to be correlated to its dynamics in the lipid bilayer and by protein–lipid interactions. These two parameters depend on the lipid internal dynamics surrounded by the lipid‐encircling discoidal scaffold protein that might differ from more unrestricted lipid bilayers observed in vesicles or cellular extracts. A solid‐state NMR spectroscopy investigation of lipid internal dynamics and thermotropism in nanodiscs is reported. The gel‐to‐fluid phase transition is almost abolished for nanodiscs, which maintain lipid fluid properties for a large temperature range. The addition of cholesterol allows fine‐tuning of the internal bilayer dynamics by increasing chain ordering. Increased site‐specific order parameters along the acyl chain reflect a higher internal ordering in nanodiscs compared with liposomes at room temperature; this is induced by the scaffold protein, which restricts lipid diffusion in the nanodisc area.

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“…Furthermore, other alternatives exist, such as bicelles for crystallization [76]-commercial kits available (in the form of lipid-detergent mixture, typically DMPC with CHAPSO); fluorinated surfactants [46] and nanodiscs (a patch of the lipid bilayer encircled by membrane scaffolding proteins) [44] for protein stabilization. Fluorinated surfactants have not gained much attention so far, probably due to its nature-it has a hydrophobic tail with the incorporation of several fluorine atoms, rendering it lipophobic and preventing effective interactions with a membrane protein, typically causing aggregation of the latter.…”

Section: Discussion and Outlookmentioning

confidence: 99%

“…For the X-ray crystallography field, it has been shown that amphipols are compatible with lipidic cubic phases [42], thus currently amphipols can only be used for in meso crystallization [43], similarly to SMALPs. There are several other approaches developed (and being actively developed) for the stabilization of membrane proteins, including nanodiscs [44], calixarens [45], fluorinated surfactants [46] and others, but they are beyond the scope of this mini-review. (From top to bottom) the concentration of detergent is increased, until the protein is extracted in the form of the complex with detergent (and some residual lipids), surrounded by free detergent-lipid micelles.…”

Section: Introductionmentioning

confidence: 99%

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An Overview of the Top Ten Detergents Used for Membrane Protein Crystallization

2017

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Abstract:To study integral membrane proteins, one has to extract them from the membrane-the step that is typically achieved by the application of detergents. In this mini-review, we summarize the top 10 detergents used for the structural analysis of membrane proteins based on the published results. The aim of this study is to provide the reader with an overview of the main properties of available detergents (critical micelle concentration (CMC) value, micelle size, etc.) and provide an idea of what detergents to may merit further study. Furthermore, we briefly discuss alternative solubilization and stabilization agents, such as polymers.

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Nanodiscs for structural and functional studies of membrane proteins

Cited by 432 publications

References 72 publications

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Extracellular vesicles as a platform for membrane‐associated therapeutic protein delivery

Lipid Internal Dynamics Probed in Nanodiscs

An Overview of the Top Ten Detergents Used for Membrane Protein Crystallization

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