Hayamizu Kenji, Koike Kota, Dodo Kosuke, Asanuma Miwako, Egami Hiromichi, Sodeoka Mikiko
Synthetic Organic Chemistry Laboratory, RIKEN Cluster for Pioneering Research 2-1 Hirosawa Wako Saitama 351-0198 Japan
RIKEN Center for Sustainable Resource Science 2-1, Hirosawa Wako Saitama 351-0198 Japan.
Chem Sci. 2023 Jul 14;14(31):8249-8254. doi: 10.1039/d2sc03112d. eCollection 2023 Aug 9.
Palladium enolates derived from β-ketocarbonyl compounds serve as key intermediates in various catalytic asymmetric reactions. We found that the palladium enolate formed from β-ketoamide is stable in air and moisture and we applied this property to develop a peptide purification system using β-ketoamide as a small affinity tag in aqueous media. A solid-supported palladium complex successfully captured β-ketoamide-tagged molecules as palladium enolates and released them in high yield upon acid treatment. Optimum conditions for the catch and release of tagged peptides from a mixture of untagged peptides were established. To demonstrate the value of this methodology in identifying the binding site of a ligand to its target protein, we purified and identified a peptide containing the ligand-binding site from the tryptic digest of cathepsin B labelled with a covalent cathepsin B inhibitor containing a β-ketoamide tag.
源自β-酮羰基化合物的钯烯醇盐在各种催化不对称反应中作为关键中间体。我们发现由β-酮酰胺形成的钯烯醇盐在空气和湿气中稳定，并且我们利用这一性质开发了一种肽纯化系统，该系统在水性介质中使用β-酮酰胺作为小的亲和标签。一种固相支持的钯配合物成功地捕获了作为钯烯醇盐的β-酮酰胺标记分子，并在酸处理后以高产率释放它们。建立了从未标记肽混合物中捕获和释放标记肽的最佳条件。为了证明该方法在鉴定配体与其靶蛋白结合位点方面的价值，我们从用含有β-酮酰胺标签的共价组织蛋白酶B抑制剂标记的组织蛋白酶B的胰蛋白酶消化物中纯化并鉴定了含有配体结合位点的肽。
