Nanodiscs as a New Tool to Examine Lipid–Protein Interactions

Denisov, Ilia G.; Schuler, Mary A.; Sligar, Stephen G.

doi:10.1007/978-1-4939-9512-7_25

Cited by 13 publications

(10 citation statements)

References 166 publications

(80 reference statements)

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“…In recent years, peptides mimicking the properties of nanodiscs, 66 i.e. embedding the membrane protein in small lipid bilayer discs surrounded by a scaffold protein, have been described.…”

Section: Membrane Protein Reconstitution In a Nutshellmentioning

confidence: 99%

Current problems and future avenues in proteoliposome research

Amati

Graf

Deutschmann

et al. 2020

Biochemical Society Transactions

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Membrane proteins (MPs) are the gatekeepers between different biological compartments separated by lipid bilayers. Being receptors, channels, transporters, or primary pumps, they fulfill a wide variety of cellular functions and their importance is reflected in the increasing number of drugs that target MPs. Functional studies of MPs within a native cellular context, however, is difficult due to the innate complexity of the densely packed membranes. Over the past decades, detergent-based extraction and purification of MPs and their reconstitution into lipid mimetic systems has been a very powerful tool to simplify the experimental system. In this review, we focus on proteoliposomes that have become an indispensable experimental system for enzymes with a vectorial function, including many of the here described energy transducing MPs. We first address long standing questions on the difficulty of successful reconstitution and controlled orientation of MPs into liposomes. A special emphasis is given on coreconstitution of several MPs into the same bilayer. Second, we discuss recent progress in the development of fluorescent dyes that offer sensitive detection with high temporal resolution. Finally, we briefly cover the use of giant unilamellar vesicles for the investigation of complex enzymatic cascades, a very promising experimental tool considering our increasing knowledge of the interplay of different cellular components.

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“…In recent years, peptides mimicking the properties of nanodiscs, 66 i.e. embedding the membrane protein in small lipid bilayer discs surrounded by a scaffold protein, have been described.…”

Section: Membrane Protein Reconstitution In a Nutshellmentioning

confidence: 99%

Current problems and future avenues in proteoliposome research

Amati

Graf

Deutschmann

et al. 2020

Biochemical Society Transactions

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“…Finally, protein-based NDs are nanometric-sized objects of discoidal shape formed by lipid bilayer patches stabilized by two encircling amphipathic helical proteins derived from apolipoprotein A1, called MSPs (Fig. 6a; for recent reviews, see Denisov and Sligar, 2017; Denisov et al ., 2019; Sligar and Denisov, 2020). The total number of MP structures established in MSP-based NDs at resolutions ⩽5 Å is 100.…”

Section: An Overview Of Surfactant Usage At the Vitrification Stepmentioning

confidence: 99%

Amphipathic environments for determining the structure of membrane proteins by single-particle electron cryo-microscopy

Bon

Michon

Popot

et al. 2021

Quart. Rev. Biophys.

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Over the past decade, the structural biology of membrane proteins (MPs) has taken a new turn thanks to epoch-making technical progress in single-particle electron cryo-microscopy (cryo-EM) as well as to improvements in sample preparation. The present analysis provides an overview of the extent and modes of usage of the various types of surfactants for cryo-EM studies. Digitonin, dodecylmaltoside, protein-based nanodiscs, lauryl maltoside-neopentyl glycol, glyco-diosgenin, and amphipols (APols) are the most popular surfactants at the vitrification step. Surfactant exchange is frequently used between MP purification and grid preparation, requiring extensive optimization each time the study of a new MP is undertaken. The variety of both the surfactants and experimental approaches used over the past few years bears witness to the need to continue developing innovative surfactants and optimizing conditions for sample preparation. The possibilities offered by novel APols for EM applications are discussed.

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“…by a membrane scaffold protein (MSP). For our cell-free screening, we first used the MSP1E3D1 nanodiscs [45][46][47] ($12-13 nm in diameter) prefilled with DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine). We purified the reaction mixtures by Ni 2+ -affinity chromatography, and we were able to identify five targets by immunoblots (8% expression rate; Tables S1 and S2).…”

Section: Cell-free Screening In Liposomesmentioning

confidence: 99%