申请人:Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V.
公开号:US10221439B2
公开(公告)日:2019-03-05
The invention relates to the in vitro and in cellulo detection of the cofactors nicotinamide adenine dinucleotide (NAD) and nicotinamide adenine dinucleotide phosphate (NADP). Provided is a sensor molecule for fluorescence- or luminescence-based detection of a nicotinamide adenine dinucleotide analyte, in particular for detecting the concentrations of NAD+, NADP+ and/or the ratios of the concentrations of NAD+/NADH and NADP+/NADPH, the sensor comprising (i) a binding protein (BP) for the nicotinamide adenine dinucleotide analyte, the BP being derived from sepiapterin reductase (SPR; EC 1.1.1.153) (ii) an SPR ligand (SPR-L) capable of intramolecular binding to said BP in the presence of the oxidized form of said analyte; and (iii) at least one fluorophore.
本发明涉及体外和细胞内检测辅因子烟酰胺腺嘌呤二核苷酸(NAD)和烟酰胺腺嘌呤二核苷酸磷酸(NADP)。本发明提供了一种基于荧光或发光的烟酰胺腺嘌呤二核苷酸分析物检测传感器分子,特别是用于检测 NAD+、NADP+ 的浓度和/或 NAD+/NADH 和 NADP+/NADPH 的浓度比,该传感器包括 (i) 用于烟酰胺腺嘌呤二核苷酸分析物的结合蛋白 (BP),该结合蛋白来源于 sepiapterin 还原酶(SPR;EC 1.1.1.153) (ii) SPR 配体 (SPR-L),能够在所述分析物的氧化形式存在时与所述 BP 进行分子内结合;以及 (iii) 至少一个荧光团。