Seitkalieva Aleksandra, Noskova Yulia, Isaeva Marina, Guzii Alla, Makarieva Tatyana N, Fedorov Sergey, Balabanova Larissa
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, Prospect 100-Letya Vladivostoka 152, 690022 Vladivostok, Russia.
Youth Research Laboratory of Recombinant DNA Technologies, Advanced Engineering School, Institute of Biotechnology, Bioengineering and Food Systems, Far Eastern Federal University, 10 Ajax Bay, Russky Island, 690922 Vladivostok, Russia.
Molecules. 2024 Dec 3;29(23):5701. doi: 10.3390/molecules29235701.
The natural 5-azaindoles, marine sponge guitarrin C and D, were observed to exert inhibitory activity against a highly active alkaline phosphatase (ALP) CmAP of the PhoA family from the marine bacterium , with IC values of 8.5 and 110 µM, respectively. The superimposition of CmAP complexes with -nitrophenyl phosphate (NPP), a commonly used chromogenic aryl substrate for ALP, and the inhibitory guitarrins C, D, and the non-inhibitory guitarrins A, B, and E revealed that the presence of a carboxyl group at C6 together with a hydroxyl group at C8 is a prerequisite for the inhibitory effect of 5-azaindoles on ALP activity. The 10-fold more active guitarrin C could compete with NPP for binding sites in the ALP active site due to similarities in size, three-dimensional structure, and the orientation of the COOH group along the phosphate group. However, the inhibition of CmAP and calf intestinal ALP (CIAP) by guitarrin C was observed to occur via a non-competitive mode of action, as evidenced by a twofold decrease in V and an unchanged K. In contrast, the kinetic model with guitarrin D, with an additional OH group at C7, reflected a mixed type of inhibition, with a decrease in both values. The sensitivity of CIAP to guitarrins C and D was shown to be slightly lower than that of CmAP, with IC values of 195 and 230 µM, respectively. Nevertheless, these findings prompted the prediction of complexes of human ALP isoenzymes with guitarrins C and D.
天然的5-氮杂吲哚类化合物，即海洋海绵吉他林C和D，被观察到对来自海洋细菌的PhoA家族的高活性碱性磷酸酶（ALP）CmAP具有抑制活性，其IC值分别为8.5和110μM。将CmAP复合物与对硝基苯磷酸酯（NPP，一种常用于ALP的生色芳基底物）以及抑制性的吉他林C、D和非抑制性的吉他林A、B和E进行叠加，结果表明C6位存在羧基以及C8位存在羟基是5-氮杂吲哚对ALP活性产生抑制作用的前提条件。活性高10倍的吉他林C由于其大小、三维结构以及COOH基团沿磷酸基团的取向与NPP相似，能够与NPP竞争ALP活性位点中的结合位点。然而，观察到吉他林C对CmAP和小牛肠碱性磷酸酶（CIAP）的抑制作用是通过非竞争性作用模式发生的，这表现为V降低两倍而K不变。相比之下，在C7位有一个额外OH基团的吉他林D的动力学模型反映出一种混合型抑制，两个值均降低。CIAP对吉他林C和D的敏感性略低于CmAP，其IC值分别为195和230μM。尽管如此，这些发现促使人们预测人ALP同工酶与吉他林C和D的复合物。
