Abstract：
Carnitine palmitoyltransferase 1A (CPT1A), which resides on the mitochondrial outer membrane, serves as the rate-limiting enzyme of fatty acid β-oxidation. Identifying the compounds targeting CPT1A warrants a promising candidate for modulating lipid metabolism. In this study, we developed a CPT1A-overexpressed mitochondrial membrane chromatography (MMC) to screen the compounds with affinity for CPT1A. Cells overexpressing CPT1A were cultured, and subsequently, their mitochondrial membrane was isolated and immobilized on amino-silica gel cross-linked by glutaraldehyde. After packing the mitochondrial membrane column, retention components of MMC were performed with LC/MS, whose analytic peaks provided structural information on compounds that might interact with mitochondrial membrane proteins. With the newly developed MMC-LC/MS approach, several Chinese traditional medicine extracts, such as Scutellariae Radix and Polygoni Cuspidati Rhizoma et Radix (PCRR), were analyzed. Five noteworthy compounds, baicalin, baicalein, wogonoside, wogonin, and resveratrol, were identified as enhancers of CPT1A enzyme activity, with resveratrol being a new agonist for CPT1A. The study suggests that MMC serves as a reliable screening system for efficiently identifying modulators targeting CPT1A from complex extracts.
摘要：
肉碱棕榈酰转移酶1A（CPT1A），位于线粒体外膜上，作为脂肪酸β-氧化的限速酶。鉴定靶向CPT1A的化合物保证了用于调节脂质代谢的有希望的候选物。在这项研究中,我们开发了CPT1A过表达的线粒体膜色谱（MMC）来筛选对CPT1A具有亲和力的化合物。培养过表达CPT1A的细胞，随后,它们的线粒体膜被分离并固定在戊二醛交联的氨基硅胶上。填充线粒体膜柱后，MMC的保留组分用LC/MS进行，其分析峰提供了可能与线粒体膜蛋白相互作用的化合物的结构信息。随着新开发的MMC-LC/MS方法，几种中药提取物，如黄芩和虎杖（PCRR），进行了分析。五个值得注意的化合物，黄芩苷,黄芩素,沃戈诺赛德,Wogonin,和白藜芦醇,被鉴定为CPT1A酶活性的增强剂，白藜芦醇是CPT1A的新激动剂。该研究表明，MMC可以作为一种可靠的筛选系统，用于从复杂提取物中有效识别靶向CPT1A的调节剂。
