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    Home > Biochemistry News > Biotechnology News > Optogenics Tools: Design Principles and Applications of OptoPB and OptoPBer.

    Optogenics Tools: Design Principles and Applications of OptoPB and OptoPBer.

    • Last Update: 2020-08-27
    • Source: Internet
    • Author: User
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    Recently, Wang Junfeng, a researcher at the Strong Magnetic Field Science Center of the Hefei Institute of Material Science of the Chinese Academy of Sciences, worked with Professors Huang Yun and Zhou Yubin of the University of Texas in the United States to develop a new type of photogenetics tool that can precisely regulate the connection between cells on a nanoscale using visible light.
    the work was published as a cover article in the Journal of the Royal Society of Chemistry (2017, 8,5275-5281), under the title Optical Control of The Age and Interorganellar Communications at Nanoscales.
    endogenous network is an important cytoscopy in the gyntic cell, which is a cavity structure consisting of a double-layer membrane.
    it interacts and exchanges information dynamically with various other membrane components within the cell (mass membrane, mitochondrial membrane, etc.), forming a huge network of endoblast membranes.
    between the endogenous mesh and the mass membrane, phospholipid inositol, which is specifically distributed on the inside of the membrane, is identified by the membrane protein on the endosomaid network, forming a membrane contact site with a distance of 10-40 nanometers.
    contact point plays a very important role in many cell physiological processes, such as membrane transport, calcium ion stabilization and lipid metabolism, and is a new research hot spot in recent years.
    but due to the lack of effective experimental tools, the study of membrane contact point formation mechanism and function is very limited.
    Photogenetics is a biological technique that uses optics and genetics to control protein activity and cellular function, and has unique advantages, such as good operability of photogenetics, real-time reversibility, precise positioning, and no trauma to cellular tissue.
    to better study the function of membrane contact sites, the two teams developed OptoPB, a photogenetic tool that uses blue light to regulate the shuttle between cytosomes and mass membranes.
    OptoPB is a fusion protein consisting mainly of photosensitive protein LOV2 and Polybasic (PB) domains.
    dark, the N-end LOV2 is like a lock, locking the C-end PB protein to the core structure.
    is exposed to blue light, the J-alpha spiral at the end of LOV2 is deconstructed and opened in seconds to release the PB protein.
    Because PB protein is rich in arginine, lysine and other alkaline amino acids can interact with acidic inositol phospholipid PI (4,5) P2 and/or PI (3,4,5) P3 on the inside of the membrane, thus anchoring the protein to the mass membrane.
    removes the light, OptoPB can quickly fall back from the membrane and disperse into the cytoste in less than a minute.
    After using MRI and molecular dynamics simulation to model the OptoPB protein under dark conditions, the team found that there are two binding pockets with negative charge on the surface of LOV2, which can well limit the PB protein with a strong positive charge to the LOV2 core domain, so as not to bind to the mass membrane.
    optoPB can be quickly transferred to the vicinity of the mass membrane under lighting conditions.
    the entire shuttle process can be reversible by alternating light and dark operation.
    In order to further regulate the formation of the membrane contact site between the endogenous mesh and the membrane membrane, the two groups worked together to insert a sequence of endogenous mesh positioning at the N end of OptoPB, forming the endogenous mesh cross-film protein OptoPBer.
    in order to precisely regulate the distance between the membrane contact points formed between the endogenous mesh and the mass membrane, a different number of (EAAAR) 4n alpha-helix sequences (n?1-8) were added between Optoper and the trans-membrane region.
    , due to the difference in the number of insertion (EAAAR) 4, the spatial distance between the endogenous mesh and the cell membrane can be effectively controlled by about 10 to 40 nanometers.
    this unique design, the two groups successfully achieved the nano-order of magnitude with blue light to remotely control the space between the endogenous mesh and membrane contact sites, and control its related functions.
    The significance of this study is that the formation of contact sites between inositol phosphate metabolism and endometrial systems of proteins can be accomplished through light, a non-invasive, highly operabable stimulus source in time and space, to achieve efficient subcellular positioning of proteins and reversible control of proteins.
    this experimental technology is free from the traditional experimental means, so that complex and difficult to regulate life activities, can be simulated under the control of blue light, has a very wide range of applications.
    , photogenetic protein engineering methods can be further extended to the study of membrane contact points between other subcellular devices.
    provides a new and effective technical means for researchers to regulate protein-membrane interaction and cell signaling path conduction between membrane and membrane.
    Wang Junfeng group postdoctoral Zhu Lei and Zhou Yubin group doctoral students He Wei, Jing Ji is the co-author of the article, Wang Junfeng, Zhou Yubin and Huang Yun as co-author of the newsletter.
    the research was funded by the National Natural Science Foundation of China, the Ministry of Science and Technology, and the China National Fund for Study Abroad, and some experiments were completed by large scientific experimental devices with steady-state strong magnetic fields.
    .
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