Bartsch Simon M, Fiedler Dorothea
Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Institut für Chemie, Humboldt-Universität zu Berlin, Berlin, Germany.
Methods Mol Biol. 2025;2972:39-49. doi: 10.1007/978-1-0716-4799-8_4.
The availability of well-defined and pure PP-InsP substrates in high quantities is pivotal to study their molecular mechanisms by means of biochemical or biophysical approaches. Here, we present a workflow employing enzymatic synthesis that allows for the quick and easy synthesis and isolation of mammalian PP-InsP isomers. With the respective enzymes, inositol pyrophosphates can be generated in a single reaction step. Isolation and purification of the desired products does not require special laboratory equipment and can be achieved by precipitation of PP-InsPs as magnesium salts. Having access to PP-InsPs will aid the field in discovering and characterizing their molecular mechanisms and binding modes.
大量获得定义明确且纯净的PP-InsP底物对于通过生化或生物物理方法研究其分子机制至关重要。在此，我们展示了一种采用酶促合成的工作流程，该流程能够快速简便地合成和分离哺乳动物PP-InsP异构体。利用相应的酶，可以在单个反应步骤中生成肌醇焦磷酸。所需产物的分离和纯化不需要特殊的实验室设备，并且可以通过将PP-InsP沉淀为镁盐来实现。能够获得PP-InsP将有助于该领域发现和表征它们的分子机制及结合模式。
