A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy

Voskoboynikova, Natalia; Karľová, Mária; Kurre, Rainer; Mulkidjanian, Armen Y.; Шайтан, К. В.; Sokolova, Olga; Steinhoff, Heinz‐Jürgen; Heinisch, Jürgen J.

doi:10.3390/jof7020118

Cited by 10 publications

(7 citation statements)

References 66 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%

“…Lipodiscs have been shown to be a valid solubilization platform for negative stain EM and low-resolution structure reconstruction (see Figure 4 C) [ 129 ] despite the prevalence of specific orientations of the protein-loaded SMALPs most commonly observed in the top views [ 171 ]. Although this method offers only modest resolution (15–20 Å), its undeniable advantage is that the structures can be obtained in a short time frame of several days [ 172 ].…”

Section: Applications Of Lipodiscs In Structural Biologymentioning

confidence: 99%

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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%

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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“…Membrane protein complexes containing photosystems I and II and light-harvesting complex have been isolated from spinach and pea thylakoids [ 151 ]. Yeast transmembrane sensor Wsc1 containing a large-soluble domain and a single transmembrane domain has been isolated, and its structural model was generated using various biophysical methods [ 152 ]. Yeast succinate dehydrogenase (Sdh4) has been isolated to study the regulation of coenzyme Q levels and how oxidative stress caused by polyunsaturated fatty acids is modulated by Cqd1 and Cqd2 proteins [ 153 ].…”

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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“…In addition to studying the above bacterial proteins, FCS in combination with nanodisc technology has been used to characterize various eukaryotic membrane proteins. Voskoboynikova et al used FCS to demonstrate successful production of monodisperse Wsc1, a yeast transmembrane cell wall stress sensor, embedded in SMALPs by SMA extraction from native S. cerevisiae cell membrane [92]. Other studies further characterized by FCS the interactions between eukaryotic membrane proteins and their substrates.…”

Section: Fcs Measurements Of Membrane Proteins Embedded In Nanodiscsmentioning

confidence: 99%

Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy

et al. 2022

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Proteins embedded in biological membranes perform essential functions in all organisms, serving as receptors, transporters, channels, cell adhesion molecules, and other supporting cellular roles. These membrane proteins comprise ~30% of all human proteins and are the targets of ~60% of FDA-approved drugs, yet their extensive characterization using established biochemical and biophysical methods has continued to be elusive due to challenges associated with the purification of these insoluble proteins. In response, the development of nanodisc techniques, such as nanolipoprotein particles (NLPs) and styrene maleic acid polymers (SMALPs), allowed membrane proteins to be expressed and isolated in solution as part of lipid bilayer rafts with defined, consistent nanometer sizes and compositions, thus enabling solution-based measurements. Fluorescence correlation spectroscopy (FCS) is a relatively simple yet powerful optical microscopy-based technique that yields quantitative biophysical information, such as diffusion kinetics and concentrations, about individual or interacting species in solution. Here, we first summarize current nanodisc techniques and FCS fundamentals. We then provide a focused review of studies that employed FCS in combination with nanodisc technology to investigate a handful of membrane proteins, including bacteriorhodopsin, bacterial division protein ZipA, bacterial membrane insertases SecYEG and YidC, Yersinia pestis type III secretion protein YopB, yeast cell wall stress sensor Wsc1, epidermal growth factor receptor (EGFR), ABC transporters, and several G protein-coupled receptors (GPCRs).

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A Three-Dimensional Model of the Yeast Transmembrane Sensor Wsc1 Obtained by SMA-Based Detergent-Free Purification and Transmission Electron Microscopy

Cited by 10 publications

References 66 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

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

Biophysical Characterization of Membrane Proteins Embedded in Nanodiscs Using Fluorescence Correlation Spectroscopy

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