Synthesis of R and S tritiated reduced β-nicotinamide adenine dinucleotide 2′ phosphate
Synthesis of R and S tritiated reduced β-nicotinamide adenine dinucleotide 2′ phosphate
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DOI：
10.1016/j.ab.2003.09.025
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发表时间：
2004-01-01
影响因子：
2.9
通讯作者：
Kohen, A
中科院分区：
文献类型：
--
作者：
McCracken, JA;Wang, T;Kohen, A
烟酰胺类化合物是一种普遍存在的辅因子，被许多生物系统用作氧化还原剂。立体特异性标记的辅因子在烟酰胺依赖性酶的许多研究中是有用的。这些辅因子的酶定向合成是相当常见的，但它们的稳定性对产率、纯度和保存提出了重大挑战。本文报道还原型R-和S-[4-H-3] β-烟酰胺腺嘌呤二核苷酸2'磷酸（NADPH）的立体专一性合成。瓦莱拉等人的方法[Biochem. Biophys. Res. Commun. 148（1987）515]修改为在2小时内产生两种同位素非对映异构体的合成程序，在纯化和冻干后产率提高至75-90%。在合成中，[4-H-3]NADP（+）作为中间体产生（如果需要，其可以被分离）。此处报告的S和R非对映异构体的比放射性分别为2.1和1.1 Ci/mmol。从无载体到痕量标记的特定放射性可以通过对程序的微小改变来实现。(C)2003爱思唯尔公司All rights reserved.
Nicotinamides are ubiquitous cofactors used by many biological systems as redox agents. Stereospecifically labeled cofactors are useful in many studies of nicotinamide-dependent enzymes. Enzyme-directed synthesis of these cofactors is rather common but their stability imposes significant challenges on yield, purity, and preservation. This paper presents the stereospecific synthesis of reduced R- and S-[4-H-3] beta-nicotinamide adenine dinucleotide 2' phosphate (NADPH). The method of Valera et al. [Biochem. Biophys. Res. Commun. 148 (1987) 515] was modified to a synthetic procedure that produces both isotopic diastereomers within 2h with an improved yield of 75-90% after purification and lyophilization. In the synthesis, [4-H-3]NADP(+) was generated as an intermediate (which can be isolated if desired). The specific radioactivities reported here are 2.1 and 1.1 Ci/mmol for the S and R diastereomers, respectively. Specific radioactivities ranging from carrier-free to trace labeling can be achieved with a minor change to the procedure. (C) 2003 Elsevier Inc. All rights reserved.