Miksic J R, Brown P R
Biochemistry. 1978 May 30;17(11):2234-8. doi: 10.1021/bi00604a034.
Reversed-phase high-pressure liquid chromatography was used to isolate acid breakdown products of reduced nicotinamide adenine dinucleotide (NADH) and products produced when NADH breakdown is catalyzed by glyceraldehyde-3-phosphate dehydrogenase (G-3-PD). Chromatographic and UV spectral data on these and related products support a mechanism for NADH acid degradation involving hydroxy addition at the nicotinamide C-6 followed by cyclization of the ring and the adjacent ribose moiety. G-3-PD is shown to catalyze a reaction in which two products are formed which are also intermediates in the acid degradation of NADH (alpha- and beta-6-hydroxynicotinamide products). Formation of the major acid products fits a three-step, first-order mechanism curve, making it possible to calculate the rate constants k2 and k3 as well as the previously determined k1.
反相高压液相色谱法用于分离还原型烟酰胺腺嘌呤二核苷酸（NADH）的酸分解产物以及由甘油醛-3-磷酸脱氢酶（G-3-PD）催化NADH分解时产生的产物。关于这些及相关产物的色谱和紫外光谱数据支持了NADH酸降解的一种机制，该机制涉及在烟酰胺C-6处进行羟基加成，随后环和相邻核糖部分发生环化。结果表明，G-3-PD催化一个反应，生成两种产物，它们也是NADH酸降解的中间体（α-和β-6-羟基烟酰胺产物）。主要酸产物的形成符合三步一级反应机制曲线，从而能够计算速率常数k2和k3以及先前测定的k1。
