de Oliveira Guilherme C, Kanamori Karina S, Auxiliadora-Martins Maria, Chini Claudia C S, Chini Eduardo N
Signal Transduction and Molecular Nutrition Laboratory, Kogod Aging Center, Department of Anesthesiology and Perioperative Medicine, Mayo Clinic College of Medicine, Rochester, USA.
Fundação de Apoio ao Ensino, Pesquisa e Assistência do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (FAEPA), São Paulo, Brazil
Bio Protoc. 2018 Jul 20;8(14). doi: 10.21769/BioProtoc.2938.
CD38 is a multifunctional enzyme involved in calcium signaling and Nicotinamide Adenine Dinucleotide (NAD) metabolism. Through its major activity, the hydrolysis of NAD, CD38 helps maintain the appropriate levels of this molecule for all NAD-dependent metabolic processes to occur. Due to current advances and studies relating NAD decline and the development of multiple age-related conditions and diseases, CD38 gained importance in both basic science and clinical settings. The discovery and development of strategies to modulate its function and, possibly, treat diseases and improve health span put CD38 under the spotlights. Therefore, a consistent and reliable method to measure its activity and explore its use in medicine is required. We describe here the methods how our group measures both the hydrolase and cyclase activity of CD38, utilizing a fluorescence-based enzymatic assay performed in a plate reader using 1,N-Ethenonicotinamide Adenine Dinucleotide (ε-NAD) and Nicotinamide Guanine Dinucleotide (NGD) as substrates, respectively.
CD38是一种参与钙信号传导和烟酰胺腺嘌呤二核苷酸（NAD）代谢的多功能酶。通过其主要活性，即NAD的水解，CD38有助于维持该分子的适当水平，以使所有依赖NAD的代谢过程得以发生。由于目前有关NAD下降与多种年龄相关病症和疾病发展的进展和研究，CD38在基础科学和临床环境中都变得至关重要。调节其功能、可能治疗疾病和改善健康寿命的策略的发现和开发使CD38成为焦点。因此，需要一种一致且可靠的方法来测量其活性并探索其在医学中的用途。我们在此描述我们小组测量CD38水解酶和环化酶活性的方法，分别使用基于荧光的酶促测定法，该测定法在酶标仪中进行，分别以1,N-乙烯基烟酰胺腺嘌呤二核苷酸（ε-NAD）和烟酰胺鸟嘌呤二核苷酸（NGD）作为底物。
