Functional incorporation of the insect odorant receptor coreceptor in tethered lipid bilayer nanoarchitectures

Kleinheinz, David; D’Onofrio, Chiara; Carraher, Colm; Ramach, Ulrich; Schuster, Bernhard; Bozdogan, Anıl; Knoll, Wolfgang; Andersson, Jakob

doi:10.1016/j.bios.2022.114024

Cited by 7 publications

(26 citation statements)

References 45 publications

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“…Prior to bilayer formation, the gold surface (functionalized with the DPhyTL/mercaptoethanol-SAM) has an ghness of 1.6 nm. 21 After bilayer formation, SthK-tBLMs had a roughness of 0.28− 0.35 nm (Figure 3A), which is in good agreement with previously reported properties of lipid bilayers. 33 AFM images of SthK-stBLMs showed features with a height of 3−4 nm (6 nm when examined with slower scanning speed (Figure 3C).…”

Section: ■ Results and Discussionsupporting

confidence: 91%

“…To first confirm bilayer formation, we imaged membranes with atomic force microscopy (AFM). Prior to bilayer formation, the gold surface (functionalized with the DPhyTL/mercaptoethanol-SAM) has an ghness of 1.6 nm . After bilayer formation, SthK-tBLMs had a roughness of 0.28–0.35 nm (Figure A), which is in good agreement with previously reported properties of lipid bilayers .…”

Section: Resultssupporting

confidence: 89%

“…Ion channel opening is indicated by a significant reduction in membrane resistance between one and four orders of magnitude. , However, as membrane resistance can also be reduced due to defect formation, we first determined the stability of the membrane system by measuring bilayer stability over time. We found no significant reductions in membrane resistance after 5 days (see Supporting Information, Figure S1) and reductions in membrane resistance can therefore confidently be attributed to ion channel function rather than defect formation.…”

Section: Resultsmentioning

confidence: 99%

“…Gold surfaces were prepared via template stripping, as described previously. 21,32 Polished silicon wafers were sonicated in acetone to remove residues from the polishing procedure, The Journal of Physical Chemistry B followed by cleaning with acidic piranha solution (3:1 sulfuric acid and a 33% hydrogen peroxide in water) for 60 min, and then rinsed thoroughly with ultrapure water and ethanol, followed by drying in a nitrogen stream. Gold Deposition.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

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Native Function of the Bacterial Ion Channel SthK in a Sparsely Tethered Lipid Bilayer Membrane Architecture

et al. 2023

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The plasma membrane protects the interiors of cells from their surroundings and also plays a critical role in communication, sensing, and nutrient import. As a result, the cell membrane and its constituents are among the most important drug targets. Studying the cell membrane and the processes it facilitates is therefore crucial, but it is a highly complex environment that is difficult to access experimentally. Various model membrane systems have been developed to provide an environment in which membrane proteins can be studied in isolation. Among them, tethered bilayer lipid membranes (tBLMs) are a promising model system providing a solvent-free membrane environment which can be prepared by self-assembly, is resistant to mechanical disturbances and has a high electrical resistance. tBLMs are therefore uniquely suitable to study ion channels and charge transport processes. However, ion channels are often large, complex, multimeric structures and their function requires a particular lipid environment. In this paper, we show that SthK, a bacterial cyclic nucleotide gated (CNG) ion channel that is strongly dependent on the surrounding lipid composition, functions normally when embedded into a sparsely tethered lipid bilayer. As SthK has been very well characterized in terms of structure and function, it is well-suited to demonstrate the utility of tethered membrane systems. A model membrane system suitable for studying CNG ion channels would be useful, as this type of ion channel performs a wide range of physiological functions in bacteria, plants, and mammals and is therefore of fundamental scientific interest as well as being highly relevant to medicine.

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Section: ■ Results and Discussionsupporting

confidence: 91%

Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

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Native Function of the Bacterial Ion Channel SthK in a Sparsely Tethered Lipid Bilayer Membrane Architecture

et al. 2023

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“…Tethered lipid bilayers − have been developed to host membrane proteins in a fluid–lipid bilayer configuration while maintaining the stability, mobility, and controllability of planar supported lipid bilayers (SLBs). − Despite these advantages, SLBs interact strongly with their substrates and are therefore often incompatible with integral membrane proteins that can bind to their substrates. − Tethered lipid bilayers take advantage of chemical tethering to separate the model lipid membrane from solid supports, so membrane-associated proteins can function without interference. − Various methods have been applied, including polymer tethers, − protein tethers, tethered bilayer lipid membranes (tBLMs) with diphytanyl ethylene glycol anchors, − and DNA–lipid conjugates, − which have been used in a variety of self-assembled membrane structures by exploiting the well-defined DNA specificity and length. This superior controllability was demonstrated in a model system using the segregation pattern of mobile tethers, and in the study of membrane fusion .…”

Section: Introductionmentioning

confidence: 99%

DNA-Tethered Lipid Membrane Formation via Solvent-Assisted Self-Assembly

Lee

Chung

2023

J. Phys. Chem. B

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DNA-tethered lipid bilayers have been used in many studies, based on the controllable and well-defined properties of DNA tethers. However, their application has been limited, because it is difficult to cover a wide range of surfaces and achieve electrical insulation. We implemented an existing method, where a DNA hybrid chip on a silica or glass surface supports a lipid membrane using solvent-assisted self-assembly. The formation of a continuous lipid bilayer was confirmed through the change in quartz crystal microbalance dissipation results, depending on the presence or absence of DNA hybrids. The fluidity of the DNA-tethered lipid membranes was analyzed using a fluorescence microscope. The electrochemical analysis demonstrated the versatility of this new technique, which can be used for sensor or electrode surface modification for biosensors or bioelectronics.

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