Characterization of lipodisc nanoparticles containing sensory rhodopsin II and its cognate transducer from Natronomonas pharaonis

Bagrov, D. V.; Voskoboynikova, Natalia; Армеев, Г. А.; Mosslehy, W.; Gluhov, Grigory; Ismagulova, T. T.; Mulkidjanian, Armen Y.; Kirpichnikov, Mikhaǐl P.; Steinhoff, Heinz‐Jürgen; Шайтан, К. В.

doi:10.1134/s0006350916060063

Cited by 8 publications

(7 citation statements)

References 45 publications

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“…At the tested particles, the height of DIBMALPs (in respect to the mica support) corresponds to the expected lipid bilayer thickness of about 3.0–4.0 nm. These data are consistent with our previously published AFM results obtained for SMALPs [ 46 ] and confirm that the DIBMA belt supports the typical lipid-bilayer thickness.…”

Section: Resultssupporting

confidence: 93%

“…Cryo-EM enables to avoid such artefacts because the rapid cryo-immobilization (plunge freezing used in this work) preserves the aqueous environment of the specimen whilst also preventing its possible damage [ 59 ]. We used AFM because of its high axial resolution that proved the native-like thickness of the assembled DIBMALPs ( Figure 3 ), in agreement with previous data obtained for SMALPs [ 46 ].…”

Section: Discussionsupporting

confidence: 86%

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Lipid Dynamics in Diisobutylene-Maleic Acid (DIBMA) Lipid Particles in Presence of Sensory Rhodopsin II

Voskoboynikova

Orekhov

Bozdaganyan

et al. 2021

IJMS

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Amphiphilic diisobutylene/maleic acid (DIBMA) copolymers extract lipid-encased membrane proteins from lipid bilayers in a detergent-free manner, yielding nanosized, discoidal DIBMA lipid particles (DIBMALPs). Depending on the DIBMA/lipid ratio, the size of DIBMALPs can be broadly varied which makes them suitable for the incorporation of proteins of different sizes. Here, we examine the influence of the DIBMALP sizes and the presence of protein on the dynamics of encased lipids. As shown by a set of biophysical methods, the stability of DIBMALPs remains unaffected at different DIBMA/lipid ratios. Coarse-grained molecular dynamics simulations confirm the formation of viable DIBMALPs with an overall size of up to 35 nm. Electron paramagnetic resonance spectroscopy of nitroxides located at the 5th, 12th or 16th carbon atom positions in phosphatidylcholine-based spin labels reveals that the dynamics of enclosed lipids are not altered by the DIBMALP size. The presence of the membrane protein sensory rhodopsin II from Natronomonas pharaonis (NpSRII) results in a slight increase in the lipid dynamics compared to empty DIBMALPs. The light-induced photocycle shows full functionality of DIBMALPs-embedded NpSRII and a significant effect of the protein-to-lipid ratio during preparation on the NpSRII dynamics. This study indicates a possible expansion of the applicability of the DIBMALP technology on studies of membrane protein–protein interaction and oligomerization in a constraining environment.

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Section: Resultssupporting

confidence: 93%

Section: Discussionsupporting

confidence: 86%

Lipid Dynamics in Diisobutylene-Maleic Acid (DIBMA) Lipid Particles in Presence of Sensory Rhodopsin II

Voskoboynikova

Orekhov

Bozdaganyan

et al. 2021

IJMS

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“…These data confirmed the presence of monodisperse nano-sized particles ( Figure 3 a). Supporting data from previous works, their intensity-weighted diameter was estimated to be in the 10-nm range [ 14 , 15 , 22 , 52 ]. More importantly, we thus demonstrated that incubation of the yeast cellular membrane fraction with the SMA copolymer in a ratio of 3:1 indeed yielded nano-sized SMALPs.…”

Section: Resultsmentioning

confidence: 96%

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

Voskoboynikova

Karľová

Kurre

et al. 2021

JoF

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The cell wall sensor Wsc1 belongs to a small family of transmembrane proteins, which are crucial to sustain cell integrity in yeast and other fungi. Wsc1 acts as a mechanosensor of the cell wall integrity (CWI) signal transduction pathway which responds to external stresses. Here we report on the purification of Wsc1 by its trapping in water-soluble polymer-stabilized lipid nanoparticles, obtained with an amphipathic styrene-maleic acid (SMA) copolymer. The latter was employed to transfer tagged sensors from their native yeast membranes into SMA/lipid particles (SMALPs), which allows their purification in a functional state, i.e., avoiding denaturation. The SMALPs composition was characterized by fluorescence correlation spectroscopy, followed by two-dimensional image acquisition from single particle transmission electron microscopy to build a three-dimensional model of the sensor. The latter confirms that Wsc1 consists of a large extracellular domain connected to a smaller intracellular part by a single transmembrane domain, which is embedded within the hydrophobic moiety of the lipid bilayer. The successful extraction of a sensor from the yeast plasma membrane by a detergent-free procedure into a native-like membrane environment provides new prospects for in vitro structural and functional studies of yeast plasma proteins which are likely to be applicable to other fungi, including plant and human pathogens.

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“…The intensity-weighted particle diameter was estimated to be in the 10 nm range consistent with previously reported data. 28,30,31,54 The DLS data indicate the monodisperse character of the SMALP preparations. Thus, incubation of NpSRII/NpHtrII 157 -liposomes with 3 : 1 SMA polymer resulted in formation of protein-containing SMA particles.…”

Section: Resultsmentioning

confidence: 98%

Characterization of an archaeal photoreceptor/transducer complex from Natronomonas pharaonis assembled within styrene–maleic acid lipid particles

et al. 2017

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Characterization of lipodisc nanoparticles containing sensory rhodopsin II and its cognate transducer from Natronomonas pharaonis

Cited by 8 publications

References 45 publications

Lipid Dynamics in Diisobutylene-Maleic Acid (DIBMA) Lipid Particles in Presence of Sensory Rhodopsin II

Lipid Dynamics in Diisobutylene-Maleic Acid (DIBMA) Lipid Particles in Presence of Sensory Rhodopsin II

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

Characterization of an archaeal photoreceptor/transducer complex from Natronomonas pharaonis assembled within styrene–maleic acid lipid particles

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