NAD⁺和FAD的8-叠氮吖啶类似物。与NAD⁺依赖性和FAD依赖性酶的合成及辅酶特性。

8-Azidoacenine analogs of NAD+ and FAD. Synthesis and coenzyme properties with NAD+-dependent and FAD-dependent enzymes.

作者信息

Koberstein R

出版信息

Eur J Biochem. 1976 Aug 1;67(1):223-9. doi: 10.1111/j.1432-1033.1976.tb10653.x.

DOI:10.1111/j.1432-1033.1976.tb10653.x

Abstract

The synthesis and purification of the 8-azidoadenine analogs of NAD+ (azido-NAD+) and FAD (AZIDO-FAD) from 8-azidoadenosine 5'-phosphate and NMN+ or FMN, respectively, is described. The coenzyme analogs are characterized by absorption, nuclear magnetic resonance and circular dichroism spectra. The two latter methods indicate a folded structure of azido-NAD+ and azido-FAD. Upon irradiation at 300 mn in aqueous solution, a change of the ultraviolet absorption spectra of the coenzyme analogs indicates photolysis of the azido group. The coenzyme properties of azido-NAD+ are demonstrated with lactate, glutamate and alcohol dehydrogenase yielding 14, 154 and 60%, respectively, of the V observed with NAD+. Concomitantly, the Km values of the coenzyme analogs are 1.7, 3.5 and 3-fold higher than those of NAD+. Azido-FAD is shown to be coenzyme of apo-glucose oxidase. The recovery of activity, however, is much slower in the presence of azido-FAD than with FAD. A final value of 66% of the activity with FAD is obtained. With apo-D-amino acid oxidase, azido-FAD is completely inactive, although it is specifically bound to the enzyme.

摘要

分别描述了由5'-磷酸-8-叠氮腺苷与NMN⁺或FMN合成和纯化NAD⁺的8-叠氮腺嘌呤类似物（叠氮-NAD⁺）和FAD的8-叠氮腺嘌呤类似物（叠氮-FAD）的过程。通过吸收光谱、核磁共振光谱和圆二色光谱对辅酶类似物进行了表征。后两种方法表明叠氮-NAD⁺和叠氮-FAD具有折叠结构。在水溶液中于300nm处照射时，辅酶类似物紫外吸收光谱的变化表明叠氮基团发生了光解。用乳酸脱氢酶、谷氨酸脱氢酶和乙醇脱氢酶证明了叠氮-NAD⁺的辅酶性质，其活性分别为NAD⁺的14%、154%和60%。同时，辅酶类似物的Km值比NAD⁺的Km值高1.7倍、3.5倍和3倍。叠氮-FAD被证明是脱辅基葡萄糖氧化酶的辅酶。然而，在有叠氮-FAD存在时，活性的恢复比有FAD时要慢得多。最终获得的活性为FAD活性的66%。对于脱辅基-D-氨基酸氧化酶，叠氮-FAD完全无活性，尽管它能特异性地与该酶结合。