Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, California 92093-0358, United States.
Bigelow Laboratory for Ocean Sciences, East Boothbay, Maine 04544, United States.
ACS Chem Biol. 2024 Jul 19;19(7):1416-1425. doi: 10.1021/acschembio.4c00169. Epub 2024 Jun 23.
Carrier proteins (CPs) play a fundamental role in the biosynthesis of fatty acids, polyketides, and non-ribosomal peptides, encompassing many medicinally and pharmacologically relevant compounds. Current approaches to analyze novel carrier-protein-dependent synthetic pathways are hampered by a lack of activity-based assays for natural product biosynthesis. To fill this gap, we turned to 3-methoxychromones, highly solvatochromic fluorescent molecules whose emission intensity and wavelength are heavily dependent on their immediate molecular environment. We have developed a solvatochromic carrier-protein-targeting probe which is able to selectively fluoresce when bound to a target carrier protein. Additionally, the probe displays distinct responses upon CP binding in carrier-protein-dependent synthases. This discerning approach demonstrates the design of solvatochromic fluorophores with the ability to identify biosynthetically active CP-enzyme interactions.
载体蛋白（CPs）在脂肪酸、聚酮和非核糖体肽的生物合成中起着至关重要的作用，包含许多具有医学和药理学相关性的化合物。目前分析新型载体蛋白依赖的合成途径的方法受到缺乏天然产物生物合成的基于活性的测定的限制。为了填补这一空白，我们转向 3-甲氧基色酮，这是一种高度溶剂化变色的荧光分子，其发射强度和波长严重依赖于其直接的分子环境。我们开发了一种溶剂化变色的载体蛋白靶向探针，当与靶载体蛋白结合时，它能够选择性地发荧光。此外，该探针在载体蛋白依赖的合成酶中与 CP 结合时表现出不同的响应。这种有区别的方法证明了设计溶剂化变色荧光团的能力，这种荧光团能够识别生物合成活性的 CP-酶相互作用。
