Assistant Professor Yohei Saito from Advanced Preventive Medical Sciences Research Center, Toya Tabei(a former graduate student at Division of Pharmaceutical Science, Graduate School of Medical Science) , Professor Kyoko Nakagawa-Goto from Faculty of Medical, Pharmaceutical and Health Sciences in Kanazawa University have developed an innovative purification strategy that utilizes boron chelates (*1) to identify intracellular target proteins of bioactive compounds.
Humans have uncovered the mysteries of biological phenomena by discovering compounds which act on the human body —bioactive compounds (*2) —that have paved the way for drug discovery. Central to this process is understanding the mechanisms through which these compounds exert their effects. A key aspect of understanding these mechanisms is identifying the specific proteins in the body that the compounds interact with, known as target proteins. One widely used method involves immobilizing a bioactive compound onto a carrier via a linker (*3), capturing the cellular protein with high affinity for the compound (the target protein), and then cleaving the linker to release the protein. However, this conventional approach faces challenges, particularly in directly analyzing the function of the target protein, due to the chemical complexity involved in linker cleavage. As a result, there is a need for more innovative methods.
The research group developed a new linker that takes advantage of the property of boroxazolidone, where the boron atom chelates with amino acids. This makes it possible to generate and dissociate boroxazolidone under mild conditions compatible with biological macromolecules (*4). Furthermore, the practicality of this system was demonstrated by showing that biocytin (*5) immobilized on a support with boroxazolidone effectively captures the target protein, avidin (*6), and that the biocytin-avidin complex dissociates from the linker simply by controlling the temperature.? The new strategy presented in this report has the potential to significantly improve the efficiency of linker cleavage, addressing a long-standing challenge. Since no special reagents are required for cleavage, target proteins can be easily eluted with simple temperature control. This approach also allows for functional analysis of the target proteins, making it an unprecedented and innovative method for target identification.
It is expected that the insights gained from this study will contribute to the identification of therapeutic targets in drug discovery and help elucidate key factors that regulate essential biological processes.
This research was published in the online edition Chemical Communications, an international journal published by the Royal Society of Chemistry in the United Kingdom, on November 8, 2024 at 9:00 a.m. (London time).
Schematic diagram of this study
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*1：Boron chelate
A chemical structure formed by the coordination bonding of a boron atom with surrounding molecules or ligands.
*2：Bioactive compounds
A compound that exhibits a specific effect by interacting with proteins, enzymes, etc. in the body of an organism, such as a drug or poison. Those that act as drugs are especially called pharmacologically active compounds.
*3：Linker
Structural part bridging between two structures, one connected with carrier and the other containing drug of interest.
*4：Biological macromolecules
Organic compounds with high molecular weight existing in cells, such as proteins, lipids and nucleic acids.
*5：Biocytin
A molecule formed from the vitamin biotin and the amino acid L-lysine. As biotin it has strong binding affinity to avidin.
*6：Avidin
A basic glycoprotein found in egg white that is the target protein of biotin.
Journal： Chemical Communications
研究者情報：斎藤 洋平
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