Detergent Alternatives: Membrane Protein Purification Using Synthetic Nanodisc Polymers

Dimitrova, Valentina S; Song, Saemee; Karagiaridi, Alexandra; Marand, Anika L.; Pinkett, Heather W.

doi:10.1007/978-1-0716-2368-8_20

Cited by 5 publications

(3 citation statements)

References 39 publications

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“…These studies revealed that proteins in SMALPs require larger energy to be released from their environment compared with MSP nanodiscs and detergent micelles, likely due to the increased stability of a protein being surrounded by its native environment, highlighting the importance of this in the study of efflux proteins. For a comprehensive review of polymer (‘native’) nanodiscs, we recommend the excellent review by Dimitrova et al [ 80 ].…”

Section: Investigating the Relationship Between Efflux Pumps And Thei...mentioning

confidence: 99%

Structural mass spectrometry approaches to understand multidrug efflux systems

Lewis

Lawrence

Hammerschmid

et al. 2023

Essays in Biochemistry

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Multidrug efflux pumps are ubiquitous across both eukaryotes and prokaryotes, and have major implications in antimicrobial and multidrug resistance. They reside within cellular membranes and have proven difficult to study owing to their hydrophobic character and relationship with their compositionally complex lipid environment. Advances in structural mass spectrometry (MS) techniques have made it possible to study these systems to elucidate critical information on their structure–function relationships. For example, MS techniques can report on protein structural dynamics, stoichiometry, connectivity, solvent accessibility, and binding interactions with ligands, lipids, and other proteins. This information proving powerful when used in conjunction with complementary structural biology methods and molecular dynamics (MD) simulations. In the present review, aimed at those not experts in MS techniques, we report on the current uses of MS in studying multidrug efflux systems, practical considerations to consider, and the future direction of the field. In the first section, we highlight the importance of studying multidrug efflux proteins, and introduce a range of different MS techniques and explain what information they yield. In the second section, we review recent studies that have utilised MS techniques to study and characterise a range of different multidrug efflux systems.

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Section: Investigating the Relationship Between Efflux Pumps And Thei...mentioning

confidence: 99%

Structural mass spectrometry approaches to understand multidrug efflux systems

Lewis

Lawrence

Hammerschmid

et al. 2023

Essays in Biochemistry

View full text Add to dashboard Cite

show abstract

“…The polymer-based purification protocol for membrane proteins in detergent-free conditions is similar to that of the detergent-based protocol ( Figure 1 ) [ 108 , 109 , 110 ]. Protease inhibitors should be included in all the purification steps as most of the target proteins are protease-sensitive.…”

Section: Detergent-free Isolation Of Membrane Proteins Using Amphipat...mentioning

confidence: 99%

Detergent-Free Isolation of Membrane Proteins and Strategies to Study Them in a Near-Native Membrane Environment

Krishnarjuna

Ramamoorthy

2022

Biomolecules

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Atomic-resolution structural studies of membrane-associated proteins and peptides in a membrane environment are important to fully understand their biological function and the roles played by them in the pathology of many diseases. However, the complexity of the cell membrane has severely limited the application of commonly used biophysical and biochemical techniques. Recent advancements in NMR spectroscopy and cryoEM approaches and the development of novel membrane mimetics have overcome some of the major challenges in this area. For example, the development of a variety of lipid-nanodiscs has enabled stable reconstitution and structural and functional studies of membrane proteins. In particular, the ability of synthetic amphipathic polymers to isolate membrane proteins directly from the cell membrane, along with the associated membrane components such as lipids, without the use of a detergent, has opened new avenues to study the structure and function of membrane proteins using a variety of biophysical and biological approaches. This review article is focused on covering the various polymers and approaches developed and their applications for the functional reconstitution and structural investigation of membrane proteins. The unique advantages and limitations of the use of synthetic polymers are also discussed.

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“…However, the situation is different when SMA nanodiscs are used, often in combination with mass spectrometry, in the study of lipid-protein interactions. The standard protocol for SMA nanodisc formation involves the joint incubation of membrane and polymer for several hours [ 47 , 48 ]. This time is many orders of magnitude longer than that of the phospholipid lateral diffusion in the plane of the membrane (diffusion coefficient of lipids is of the order of 10 −7 cm 2 s −1 ) [ 49 , 50 ].…”

Section: Birth and Perpetuation Of A Mythmentioning

confidence: 99%

The Myth of The Annular Lipids

Gómez‐Fernández

Goñi

2022

Biomedicines

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In the early 1970s, the existence of a “lipid annulus” stably surrounding the individual intrinsic protein molecules was proposed by several authors. They referred to a number of lipid molecules in slow exchange with the bulk lipid in the bilayer, i.e., more or less protein-bound, and more ordered than the bulk lipid. The annular lipids would control enzyme activity. This idea was uncritically accepted by most scientists working with intrinsic membrane proteins at the time, so that the idea operated like a myth in the field. However, in the following decade, hard spectroscopic and biochemical evidence showed that the proposed annular lipids were not immobilized for a sufficiently long time to influence enzyme or transporter activity, nor were they ordered by the protein. Surprisingly, forty years later, the myth survives, and the term ‘annular lipid’ is still in use, in a different, but even more illogical sense.

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Detergent Alternatives: Membrane Protein Purification Using Synthetic Nanodisc Polymers

Cited by 5 publications

References 39 publications

Structural mass spectrometry approaches to understand multidrug efflux systems

Structural mass spectrometry approaches to understand multidrug efflux systems

Detergent-Free Isolation of Membrane Proteins and Strategies to Study Them in a Near-Native Membrane Environment

The Myth of The Annular Lipids

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