Garavaglia Silvia, Galizzi Alessandro, Rizzi Menico
DISCAFF-INFM, University of Piemonte Orientale Amedeo Avogadro, 28100 Novara, Italy.
J Bacteriol. 2003 Aug;185(16):4844-50. doi: 10.1128/JB.185.16.4844-4850.2003.
NADP is essential for biosynthetic pathways, energy, and signal transduction. In living organisms, NADP biosynthesis proceeds through the phosphorylation of NAD with a reaction catalyzed by NAD kinase. We expressed, purified, and characterized Bacillus subtilis NAD kinase. This enzyme represents a new member of the inorganic polyphosphate [poly(P)]/ATP NAD kinase subfamily, as it can use poly(P), ATP, or other nucleoside triphosphates as phosphoryl donors. NAD kinase showed marked positive cooperativity for the substrates ATP and poly(P) and was inhibited by its product, NADP, suggesting that the enzyme plays a major regulatory role in NADP biosynthesis. We discovered that quinolinic acid, a central metabolite in NAD(P) biosynthesis, behaved like a strong allosteric activator for the enzyme. Therefore, we propose that NAD kinase is a key enzyme for both NADP metabolism and quinolinic acid metabolism.
烟酰胺腺嘌呤二核苷酸磷酸(NADP)对于生物合成途径、能量代谢及信号转导至关重要。在活生物体中,NADP的生物合成是通过烟酰胺腺嘌呤二核苷酸(NAD)的磷酸化反应进行的,该反应由NAD激酶催化。我们对枯草芽孢杆菌NAD激酶进行了表达、纯化及特性鉴定。这种酶代表了无机多聚磷酸[poly(P)]/ATP NAD激酶亚家族的一个新成员,因为它可以使用多聚磷酸、ATP或其他核苷三磷酸作为磷酰基供体。NAD激酶对底物ATP和多聚磷酸表现出明显的正协同性,并受到其产物NADP的抑制,这表明该酶在NADP生物合成中起主要调节作用。我们发现,喹啉酸作为NAD(P)生物合成中的一种核心代谢物,对该酶表现出强效别构激活剂的作用。因此,我们提出NAD激酶是NADP代谢和喹啉酸代谢的关键酶。