JP-2026074658-A - Ion channel protein evaluation model, ion channel current measuring device, and method for manufacturing the ion channel protein evaluation model

Abstract

[Problem] To provide an ion channel protein evaluation model and a method for producing the same, which can incorporate multiple channel proteins into a single lipid bilayer. [Solution] An ion channel protein evaluation model in which an ion channel protein is provided penetrating a lipid bilayer, and the ion channel protein is linked to gel beads. [Selection Diagram] Figure 6

Inventors

• 井出 徹
• 平野 美奈子
• 朝倉 真実

Assignees

• 国立大学法人 岡山大学

Dates

Publication Date

20260507

Application Date

20241021

Claims (6)

1. An ion channel protein evaluation model in which ion channel proteins are arranged across a lipid bilayer, and these ion channel proteins are linked to gel beads.
2. An ion channel current measuring device comprising the ion channel protein evaluation model described in claim 1.
3. The ion channel current measuring device according to claim 2, comprising one or more suction ports for aspirating and fixing one or more gel beads, a space for holding the one or more gel beads, and a space for holding a water-soluble solvent on the side opposite to the one or more suction ports.
4. A step of bringing ion channel proteins immobilized on gel beads into contact with the surface of a lipid dissolution layer laminated on a water-soluble solvent layer, and A method for producing an ion channel protein evaluation model according to claim 1, comprising the step of inserting the ion channel protein to the interface between the water-soluble solvent layer and the lipid-soluble solution layer.
5. The manufacturing method according to claim 4, wherein the ion channel protein is contained within a membrane vesicle linked to the gel beads.
6. The manufacturing method according to claim 5, wherein the membrane vesicles are linked to the gel beads by a His-tag or Biotin-tag.

Description

Application for application of Article 30, Paragraph 2 of the Patent Law (Part 1) Date of publication of the website October 23, 2023 Website address https://www2.aeplan.co.jp/bsj2023/index.html https://www2.aeplan.co.jp/bsj2023/processings.html (Part 2) Dates of the event November 14 to November 16, 2023 Name of the meeting and venue The 61st Annual Meeting of the Biophysical Society of Japan Nagoya Congress Center (1-1 Atsuta-Nishimachi, Atsuta-ku, Nagoya, Aichi) (Part 3) Date of publication of the website June 21, 2024 Website address https://www. c-linkage.co.jp/iupab2024-bsj-kyoto/ https://www.c-linkage.co.jp/iupab2024-bsj-kyoto/files/online-abstracts. PDF (Part 4) Dates: June 24, 2024 to June 28, 2024 Name of the meeting, location: 21st IUPAB Congress 2024 (IUPAB2024) Kyoto International Conference Center (422 Iwakura Osagi-cho, Sakyo-ku, Kyoto City, Kyoto Prefecture) (Part 5) Website posting dates: July 8, 2024 to July 31, 2024 Website address: https://www.c-linkage.co.jp/iupab2024-bsj-kyoto/ https://www.c-linkage.co.jp jp/iupab2024-bsj-kyoto/files/poster-sessions26. pdf This invention relates to an ion channel protein evaluation model, an ion channel current measuring device, and a method for manufacturing the ion channel protein evaluation model. Ion channel proteins are transmembrane proteins that exist across the cell's biological membrane (lipid bilayer). They play a crucial role in various signal transduction processes within and outside the cell by regulating the permeability of ions. They are extremely important in living organisms. It is known that abnormalities in ion channel proteins (hereinafter also referred to as "channel proteins" in this specification) often induce serious diseases. For example, muscular dystrophy develops when there is an abnormality in calcium channels, alternating hemiplegia in children develops when there is an abnormality in Na + /K + - countertransport ATPase, cystic fibrosis develops when there is an abnormality in chloride ion channels, and periodic disorder develops when there is an abnormality in voltage-opening potassium channels. These diseases are known as "channel diseases," and the development of effective treatments is urgently needed. Of the 6,650 human genes that could potentially be targeted by drugs, approximately 15% are said to be channel proteins, highlighting the need for highly efficient and accurate screening methods. One in vitro screening method for channel disease treatments involves creating an artificial biological membrane (lipid bilayer), incorporating channel proteins, and measuring the electrical current. While this method has many advantages, such as being applicable to various cell types, being suitable for intracellular membrane channels, and offering a high degree of flexibility in the measurement environment, it suffers from low efficiency. The conventional method for measuring the current of such channel proteins involves artificially forming a lipid bilayer in aqueous solution, inserting the ion channel protein into the lipid bilayer via membrane fusion, and then measuring the ionic current. However, this method requires a considerable amount of time for the artificial formation of the lipid bilayer in aqueous solution. Furthermore, the process of inserting the ion channel protein into the formed lipid bilayer also requires a significant amount of time, and it is difficult to insert multiple ion channel proteins simultaneously, making it extremely inefficient. Therefore, a method has been proposed in which channel proteins are bound to gold electrodes coated with a hydrophilic membrane and inserted into an artificially fabricated lipid bilayer (Patent Document 1). While this method can shorten the time required for lipid bilayer formation, there are still obstacles to inserting multiple ion channel proteins, making multi-channelization practically impossible. Therefore, a more efficient ion channel protein evaluation model capable of incorporating multiple ion channel proteins into a lipid bilayer is needed. Patent No. 6632826 Schematic diagram of lipids and lipid bilayers.An explanatory diagram of the method for producing the ion channel protein evaluation model of the present invention.An explanatory diagram of the method for producing the ion channel protein evaluation model of the present invention.Schematic diagram of ion channel proteins linked to gel beads.Schematic diagram of a membrane vesicle.A schematic diagram of an example of the ion channel current measuring device of the present invention.A schematic diagram for fabricating the ion channel current measuring device of the present invention.Measurement results for Example 1.Measurement results for Example 2.Measurement results for Example 3.Measurement results for Example 4.Measurement results for Examples 5-8.An example of an ion channel current measuring device and test results.Examples of other embodiments of an ion channel current measuring device and test res