State Key Laboratory of Membrane Biology, College of Future Technology, Institute of Molecular Medicine, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing, 100871, China; National Biomedical Imaging Center, Peking University, Beijing, 100871, China.
State Key Laboratory of Membrane Biology, College of Future Technology, Institute of Molecular Medicine, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing, 100871, China; National Biomedical Imaging Center, Peking University, Beijing, 100871, China; Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, 100871, China.
Biochem Biophys Res Commun. 2022 Sep 17;621:168-175. doi: 10.1016/j.bbrc.2022.06.100. Epub 2022 Jul 4.
Nicotinic acid adenine dinucleotide phosphate (NAADP) is a signaling molecule that can induce calcium release from intracellular acidic stores. However, proteins that bind to NAADP are understudied. Here, we identify aspartate dehydrogenase domain-containing protein (ASPDH) as an NAADP-binding protein through biochemical purification from pig livers. Isothermal titration calorimetry (ITC) experiment using the recombinantly expressed protein shows a 1:1 binding stoichiometry and a Kd of 455 nM between NAADP and mouse ASPDH. In contrast, recombinantly expressed Jupiter microtubule-associated homolog 2 (JPT2) and SM-like protein LSM12, two proteins previously identified as NAADP-receptors, show no binding in ITC experiments.
烟酰胺腺嘌呤二核苷酸磷酸（NAADP）是一种信号分子，可以从细胞内酸性储存库中诱导钙释放。然而，与 NAADP 结合的蛋白质研究较少。在这里，我们通过从猪肝脏中进行生化纯化，鉴定出天冬氨酸脱氢酶结构域蛋白（ASPDH）为 NAADP 结合蛋白。使用重组表达蛋白进行的等温滴定量热法（ITC）实验表明，NAADP 与小鼠 ASPDH 之间存在 1:1 的结合比例和 455 nM 的 Kd。相比之下，先前鉴定为 NAADP 受体的重组表达木星微管相关同源物 2（JPT2）和 SM 样蛋白 LSM12 在 ITC 实验中没有显示出结合。
