Goulas P
Département des Sciences Naturelles, Faculté des Sciences, Pau.
Eur J Biochem. 1987 Oct 15;168(2):469-73. doi: 10.1111/j.1432-1033.1987.tb13440.x.
Examination of the model of the fixation site of the adenosine phosphate part of NAD+ on horse liver alcohol dehydrogenase led us to synthesize a NAD+ analogue N6-[N-(8-amino-3,6-dioxaoctyl)carbamoylmethyl]-NAD+ in order to alkylate the carboxylic acid group of Asp-273 and to convert the normally dissociable coenzyme into a permanently bound prosthetic group. This NAD+ analogue is coupled to the horse liver alcohol dehydrogenase in the ternary complex formed with pyrazole. In these conditions the degree of fixation varies between 0.4 and 0.58 coenzyme molecule/enzyme subunit molecule. The N6-[N-(8-amino-3,6-dioxaoctyl)carbamoylmethyl]NAD+ acts as a true prosthetic group which can be reduced and reoxidized by a coupled substrate reaction and the internal activity of this holoenzyme corresponds to the amount of analogue incorporated.
对烟酰胺腺嘌呤二核苷酸（NAD⁺）磷酸腺苷部分在马肝醇脱氢酶上的结合位点模型进行研究后，我们合成了一种NAD⁺类似物N6-[N-(8-氨基-3,6-二氧杂辛基)氨甲酰甲基]-NAD⁺，以便使天冬氨酸-273的羧基烷基化，并将通常可解离的辅酶转化为永久结合的辅基。这种NAD⁺类似物在与吡唑形成的三元复合物中与马肝醇脱氢酶偶联。在这些条件下，结合程度在0.4至0.58个辅酶分子/酶亚基分子之间变化。N6-[N-(8-氨基-3,6-二氧杂辛基)氨甲酰甲基]NAD⁺作为一种真正的辅基，可通过偶联底物反应进行还原和再氧化，并且这种全酶的内在活性与掺入的类似物量相对应。
