Gavrilova E M, Kiseleva N I, Egorov A M, Berezin I V
Biokhimiia. 1981 Feb;46(2):306-13.
A method resulting in ATP-insulin conjugates by covalent binding of ATP modified at C(6) amino group of the adenine residue with insulin was developed. The modified ATP was bound to insulin by means of metha-p-toluene sulfonate-N-cyclohezyl Nf [2-morpholinyl(4)ethyl]-carbodiimide. The ATP analogs and ATP-insulin conjugates possess the coenzyme activity in a reaction of luciferin oxidation by luciferase from the fireflies Luciola mingrelica. the catalytic properties of soluble and immobilize on CNBR-activated. Sepharose enzymes in reactions with native ATR, its modified derivatives and ATP--insulin conjugates were compared. The bioluminescence reaction involving ATP--insulin conjugate is inhibited by antibodies against insulin. This effect can form a basis for insulin detection in solution, which is based on competitive binding of free and antibody-labelled ATP--insulin conjugates.
开发了一种通过将腺嘌呤残基的C(6)氨基修饰的ATP与胰岛素共价结合来生成ATP-胰岛素缀合物的方法。修饰后的ATP通过对甲苯磺酸-N-环己基N'-[2-吗啉基(4)乙基]-碳二亚胺与胰岛素结合。ATP类似物和ATP-胰岛素缀合物在萤火虫明纹暗萤的荧光素酶氧化荧光素的反应中具有辅酶活性。比较了可溶性和固定在CNBR活化的琼脂糖上的酶在与天然ATP、其修饰衍生物和ATP-胰岛素缀合物反应中的催化特性。涉及ATP-胰岛素缀合物的生物发光反应被抗胰岛素抗体抑制。这种效应可以为溶液中胰岛素的检测奠定基础,该检测基于游离和抗体标记的ATP-胰岛素缀合物的竞争性结合。