Construction and Structural Analysis of Tethered Lipid Bilayer Containing Photosynthetic Antenna Proteins for Functional Analysis

Sumino, Ayumi; Dewa, Takehisa; Takeuchi, Toshikazu; Sugiura, Ryuta; Sasaki, Nobuaki; Misawa, Nobuo; Tero, Ryugo; Urisu, Tsuneo; Gardiner, Alastair T.; Cogdell, Richard J.; Hashimoto, Hideki; Nango, Mamoru

doi:10.1021/bm200585y

Cited by 35 publications

(31 citation statements)

References 61 publications

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“…The third process of the adsorbed vesicles on substrates is an adhesive vesicular layer that stably forms, and the transformation to a planar membrane does not proceed (Figure 1c) [59,63]. Once the vesicular layer forms, it rarely turns to a planar membrane spontaneously, but there are several known ways to stimulate adhesive vesicles to rupture: addition of a reagent working as a membrane fuser (Ca 2+ [58], poly-ethylene glycol [68]) or an amphipathic viral peptide [69], osmotic pressure [60], mild sonication [70] and freeze-and-thaw [71]. …”

Section: Substrate Effects On Slb Formation Processmentioning

confidence: 99%

“…Generally used linker molecules are polymers covalently attached to the bilayer and the substrate, or avidin attached on the substrate binding with biotinyl lipids doped in the lipid bilayer. Dewa et al extensively studied the diffusion properties of bacterial photosynthetic membrane proteins, light-harvesting complex 2 (LH2) and light-harvesting core complex (LH1-RC), in SLBs and tethered bilayers by FRAP [70,118,119]. The tethered bilayer of DOPC containing 1% biotinyl dioleoylphosphatidylethanolamine (N-biotinyl-DOPE) is prepared on an avidin-modified glass cover slip [120,121].…”

Section: Substrate Effects On the Molecular Diffusion In Slbmentioning

confidence: 99%

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Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

2012

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Supported lipid bilayers are artificial lipid bilayer membranes existing at the interface between solid substrates and aqueous solution. Surface structures and properties of the solid substrates affect the formation process, fluidity, two-dimensional structure and chemical activity of supported lipid bilayers, through the 1–2 nm thick water layer between the substrate and bilayer membrane. Even on SiO2/Si and mica surfaces, which are flat and biologically inert, and most widely used as the substrates for the supported lipid bilayers, cause differences in the structure and properties of the supported membranes. In this review, I summarize several examples of the effects of substrate structures and properties on an atomic and nanometer scales on the solid-supported lipid bilayers, including our recent reports.

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Section: Substrate Effects On Slb Formation Processmentioning

confidence: 99%

Section: Substrate Effects On the Molecular Diffusion In Slbmentioning

confidence: 99%

Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

2012

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“…Systems have been reported in which bilayers are decoupled from solid supports by ultrathin, hydrated polymer cushions [108–111] or short linear tethers of hydrophilic oligomers [112–114]. Both tethering approaches lead to the stabilization of well-defined hydrated cushion layers between membrane and carrier substrate [113], important to maintain in-plane fluidity of the bilayers, to protect the protein from denaturation or aggregation following unspecific adsorption to the otherwise bare substrate surface and to permit the incorporation of transmembrane proteins [115–117]. A system developed in the Lösche group [113], the sparsely-tethered bilayer lipid membrane (stBLM), is widely applicable for the biophysical characterization of lipid-protein interactions and is particularly useful for high-resolution neutron scattering investigations [115, 118].…”

Section: 4 Structure Of Membrane-bound Ptenmentioning

confidence: 99%

PtdIns(4,5)P2-Mediated Cell Signaling: Emerging Principles and PTEN as a Paradigm for Regulatory Mechanism

Gericke

Leslie

Lösche

et al. 2013

Advances in Experimental Medicine and Biology

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PI(4,5)P2 (phosphatidylinositol 4,5-bisphosphate) is a relatively common anionic lipid that regulates cellular functions by multiple mechanisms. Hydrolysis of PI(4,5)P2 by phospholipase C yields inositol trisphosphate and diacylglycerol. Phosphorylation by phosphoinositide 3-kinase yields PI(3,4,5)P3, which is a potent signal for survival and proliferation. Also, PI(4,5)P2 can bind directly to integral and peripheral membrane proteins. As an example of regulation by PI(4,5)P2, we discuss phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in detail. PTEN is an important tumor suppressor and hydrolyzes PI(3,4,5)P3. PI(4,5)P2 enhances PTEN association with the plasma membrane and activates its phosphatase activity. This is a critical regulatory mechanism, but a detailed description of this process from a structural point of view is lacking. The disordered lipid bilayer environment hinders structural determinations of membrane-bound PTEN. A new method to analyze membrane-bound protein measures neutron reflectivity for proteins bound to tethered phospholipid membranes. These methods allow determination of the orientation and shape of membrane-bound proteins. In combination with molecular dynamics simulations, these studies will provide crucial structural information that can serve as a foundation for our understanding of PTEN regulation in normal and pathological processes.

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“…This type of bilayer has the disadvantage that the interaction between one lipid monolayer and the support is very strong, possibly perturbing the physicochemical properties. Thus, several groups have taken to producing tethered bilayers in which the lipids are attached to the support, leaving an aqueous space between the membrane and the support (Sumino et al 2011). This is achieved by attaching the proteins and/or lipids to the surface with spacers.…”

Section: Variations On a Theme: Supported Lipid Bilayers And Giant Unmentioning

confidence: 99%

Building Model Membranes with Lipids and Proteins: Dangers and Challenges

Sturgis

2014

Membrane Proteins Production for Structural Analysis

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First, it is necessary to understand why biochemists wish to reconstitute membrane proteins into lipid vesicles and nonnative systems, and indeed there are several different reasons why this path could be taken. It is important to realize that there can be multiple objectives, as the shortcomings of a particular method for one objective might be an advantage for another. Historically, reconstitution has been aimed at both structural and functional studies. Structural studies can include not only the obvious determination of structure but also more subtle aspects of structure and the interplay of environment and structure, or the dynamics of the structure. Similarly, there are different aspects of function that can be targeted, many of which are accessible only in reconstituted systems.Structural studies, such as high-resolution three-dimensional (3D) structural determination, can be conducted using 2D crystals formed in lipid membranes by using electron diffraction (e.g., Wang and Kühlbrandt 1992); however, these structures rarely compete with the structures obtained from X-ray diffraction (XRD) of 3D crystals. However, this approach can be particularly rewarding when the organization of lipids around a membrane protein is of interest. For example, initial studies of aquaporins by X-ray crystallography did not find any associated lipids (Murata et al. 2000); however, 2D crystals of aquaporin can be formed in several different lipids, and indeed in the structure obtained by electron diffraction (Gonen et al. 2005), many lipids were resolved in the crystal structure, including a few that were not in direct contact with the protein. Reconstitution for such structural studies, which can provide high quality 3D structures and information on the immediate environment, this is one possible reason to make artificial membranes. The objective is high protein density, and so reconstitutions are typically at lipid-to-protein ratio of less than 0.5 mg/mg.For functional studies, reconstitution is often necessary because membrane proteins have transport activities. So ion channels are typically reconstituted into lipid bilayers for electrophysiological or transport measurements; for example, the KcsA I. Mus-Veteau (ed.), Membrane Proteins Production for Structural Analysis,

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Construction and Structural Analysis of Tethered Lipid Bilayer Containing Photosynthetic Antenna Proteins for Functional Analysis

Cited by 35 publications

References 61 publications

Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

PtdIns(4,5)P2-Mediated Cell Signaling: Emerging Principles and PTEN as a Paradigm for Regulatory Mechanism

Building Model Membranes with Lipids and Proteins: Dangers and Challenges

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