Torreilles J, Guérin M C, Descomps B
Biochimie. 1986 Apr;68(4):565-8. doi: 10.1016/s0300-9084(86)80200-0.
We describe a procedure using immobilized nicotinamide as an affinity chromatographic ligand for the binding of NAD(P)+-dependent dehydrogenases. The procedure involves preparation of nicotinamide N1-(N-(6-aminohexyl)-acetamide)-agarose and modification of the immobilized nicotinamide by the addition of a ketone or an aldehyde to form an adduct. The nicotinamide, which has no affinity for dehydrogenase, becomes a very specific ligand of dehydrogenase, which binds the ketone or the aldehyde as substrate or inhibitor. In tests, the adduct prepared with immobilized nicotinamide and sodium pyruvate bound specifically to lactate dehydrogenase (EC 1.1.1.27), whereas the adduct prepared with alpha-ketoglutarate bound to glutamate dehydrogenase (EC 1.4.1.3). This technique enables the rapid isolation of a given dehydrogenase.
我们描述了一种使用固定化烟酰胺作为亲和色谱配体来结合NAD(P)+依赖性脱氢酶的方法。该方法包括制备烟酰胺N1-(N-(6-氨基己基)-乙酰胺)-琼脂糖，并通过添加酮或醛来修饰固定化烟酰胺以形成加合物。原本对脱氢酶没有亲和力的烟酰胺，变成了脱氢酶的一种非常特异性的配体，它将酮或醛作为底物或抑制剂结合。在测试中，用固定化烟酰胺和丙酮酸钠制备的加合物特异性地结合乳酸脱氢酶（EC 1.1.1.27），而用α-酮戊二酸制备的加合物则结合谷氨酸脱氢酶（EC 1.4.1.3）。这项技术能够快速分离特定的脱氢酶。
