Knowlan Kevin, Hoop Cody L, Tarasova Nadya I
Cancer Innovation Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, P.O. Box B, Frederick, MD, 21702, USA.
Anal Biochem. 2025 Jul 11;706:115944. doi: 10.1016/j.ab.2025.115944.
Cis-aconitate decarboxylase (ACOD1) is a key enzyme converting cis-aconitate to itaconate, which has therapeutic potential for inflammatory diseases. Existing methods to measure ACOD1 activity and itaconate are often expensive and complex. We developed a novel, high-throughput spectrophotometric assay using the Fürth-Herrmann reaction. Our method quantifies ACOD1-catalyzed itaconate production by leveraging distinct absorbance ratios of cis-aconitate and itaconate at 386 nm and 440 nm. We optimized parameters, characterized human ACOD1 kinetics, and determined an IC for citraconate consistent with previous reports. This simple, fast, and reliable assay, requiring only a UV-Vis spectrophotometer, will accelerate screening for ACOD1 modulators, speeding up therapeutic development.
顺乌头酸脱羧酶（ACOD1）是一种将顺乌头酸转化为衣康酸的关键酶，衣康酸对炎症性疾病具有治疗潜力。现有的测量ACOD1活性和衣康酸的方法通常昂贵且复杂。我们开发了一种使用菲尔特 - 赫尔曼反应的新型高通量分光光度法。我们的方法通过利用顺乌头酸和衣康酸在386纳米和440纳米处不同的吸光度比值来定量ACOD1催化产生的衣康酸。我们优化了参数，表征了人类ACOD1的动力学，并确定了与先前报道一致的柠康酸的IC。这种简单、快速且可靠的检测方法仅需一台紫外可见分光光度计，将加速对ACOD1调节剂的筛选，加快治疗药物的研发进程。
