NMR Structural and Dynamical Investigation of the Isolated Voltage-Sensing Domain of the Potassium Channel KvAP: Implications for Voltage Gating

Шенкарев, Захар О.; Paramonov, Alexander S.; Lyukmanova, Ekaterina N.; Shingarova, L. N.; Yakimov, S. A.; Dubinnyi, Maxim A.; Chupin, Vladimir; Kirpichnikov, Mikhaǐl P.; Blommers, Marcel J. J.; Arseniev, Alexander S.

doi:10.1021/ja909752r

Cited by 62 publications

(70 citation statements)

References 58 publications

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“…Included in this collection are integral membrane proteins, such as G-protein coupled receptors (GPCRs) (30–35), ion channels (36–39), transporters (40–45), pores and toxins (46, 47), receptors (48–51), cytochrome c oxidase (52); each incorporated into Nanodiscs with precisely controlled compositions. A pictorial "Rogues Gallery" of these embedded proteins is represented in Figure 2.…”

Section: Nanodiscs For Analysis Of Lipid-protein Interactionsmentioning

confidence: 99%

Nanodiscs as a New Tool to Examine Lipid–Protein Interactions

Schuler

Denisov

Sligar

2012

Methods in Molecular Biology

139

131

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Nanodiscs are self-assembled discoidal fragments of lipid bilayers 8 – 16 nm in diameter, stabilized in solution by amphipathic helical scaffold protein. As stable and highly soluble membrane mimetics with controlled lipid composition and ability to add affinity tags to the scaffold protein, Nanodiscs represent an attractive model system for solubilization, isolation, purification, and biophysical and biochemical studies of membrane proteins. We overview various approaches to the structural and functional studies of different classes of integral membrane proteins such as ion channels, transporters, GPCR and other receptors, membrane enzymes, blood coagulation cascade proteins, et cetera incorporated into Nanodiscs with the special focus on the advantages provided by homogeneity, ability to control oligomerization state of the target protein and lipid composition of the bilayer. Special attention is paid to the opportunities provided by Nanodisc system for the detailed studies of the role of different lipid properties and protein – lipid interactions in the functional behavior of membrane proteins.

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Section: Nanodiscs For Analysis Of Lipid-protein Interactionsmentioning

confidence: 99%

Nanodiscs as a New Tool to Examine Lipid–Protein Interactions

Schuler

Denisov

Sligar

2012

Methods in Molecular Biology

139

131

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“…Detergent micelles can retain the proper tertiary structure of some MPs [4][5][6][7] and have relatively small size to provide the NMR spectra with narrow lines; detergents are cheap and some of them are commercially available in a perdeuterated form. On the other hand, micelles have spherical shape and do not contain any patch of a planar lipid bilayer, which can severely affect the spatial structure of MPs, especially of those, containing juxtamembrane regions that tend to interact with the membrane surface 8 .…”

Section: Introductionmentioning

confidence: 99%

Characterization of Small Isotropic Bicelles with Various Compositions

et al. 2016

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Structural studies of membrane proteins are of great importance and interest, with solution and solid state NMR spectroscopy being very promising tools for that task. However, such investigations are hindered by a number of obstacles, and in the first place by the fact that membrane proteins need an adequate environment that models the cell membrane. One of the most widely used and prospective membrane mimetics is isotropic bicelles. While large anisotropic bicelles are well-studied, the field of small bicelles contains a lot of "white spots". The present work reports the radii of particles and concentration of the detergents in the monomeric state in solutions of isotropic bicelles, formed by 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propanesulfonate (CHAPSO), and sodium cholate, as a function of lipid/detergent ratio and temperature. These parameters were measured using (1)H NMR diffusion spectroscopy for the bicelles composed of lipids with saturated fatty chains of different length and lipids, containing unsaturated fatty acid residue. The influence of a model transmembrane protein (membrane domain of rat TrkA) on the properties of bicelles and the effect of the bicelle size and composition on the properties of the transmembrane protein were investigated with heteronuclear NMR and nuclear Overhauser effect spectroscopy. We show that isotropic bicelles that are applicable for solution NMR spectroscopy behave as predicted by the theoretical models and are likely to be bicelles rather than mixed micelles. Using the obtained data, we propose a simple approach to control the size of bicelles at low concentrations. On the basis of our results, we compared different rim-forming agents and selected CHAPS as a detergent of choice for structural studies in bicelles, if the deuteration of the detergent is not required.

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“…The first had the VSD solubilized in DPC/LDAO mixed micelles (50 kDa complex at 42 C) (Shenkarev et al 2010a) and provided structural information that matched well with the crystal structure of the isolated VSD in n-octyl-b-D-glucoside (Jiang et al 2003), which has four transmembrane a-helical segments (S1-S4); it was concluded that this NMR structure of the VSD is in a conformation that corresponds to the open-state of the KvAP channel. The second study determined the structure of the VSD in D7PC (1,2-diheptanoyl-sn-glycero-3-phosphocholine) micelles (50-60 kDa complex at 45 C) (Butterwick and MacKinnon 2010) ( Figure 11C) and was generally in overall agreement with the earlier study and with the crystal structure of the isolated VSD.…”

Section: Voltage-sensor Domain Of the Potassium Channel Kvapmentioning

confidence: 83%

NMR structures of polytopic integral membrane proteins

Patching

2011

Molecular Membrane Biology

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Membrane proteins represent up to 30% of the proteins in all organisms, they are involved in many biological processes and are the molecular targets for around 50% of validated drugs. Despite this, membrane proteins represent less than 1% of all high-resolution protein structures due to various challenges associated with applying the main biophysical techniques used for protein structure determination. Recent years have seen an explosion in the number of high-resolution structures of membrane proteins determined by NMR spectroscopy, especially for those with multiple transmembrane-spanning segments. This is a review of the structures of polytopic integral membrane proteins determined by NMR spectroscopy up to the end of the year 2010, which includes both β-barrel and α-helical proteins from a number of different organisms and with a range in types of function. It also considers the challenges associated with performing structural studies by NMR spectroscopy on membrane proteins and how some of these have been overcome, along with its exciting potential for contributing new knowledge about the molecular mechanisms of membrane proteins, their roles in human disease, and for assisting drug design.

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NMR Structural and Dynamical Investigation of the Isolated Voltage-Sensing Domain of the Potassium Channel KvAP: Implications for Voltage Gating

Cited by 62 publications

References 58 publications

Nanodiscs as a New Tool to Examine Lipid–Protein Interactions

Nanodiscs as a New Tool to Examine Lipid–Protein Interactions

Characterization of Small Isotropic Bicelles with Various Compositions

NMR structures of polytopic integral membrane proteins

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