Cytidine Diphosphoramidate Kinase: An Enzyme Required for the Biosynthesis of the O-Methyl Phosphoramidate Modification in the Capsular Polysaccharides of Campylobacter jejuni.
Cytidine Diphosphoramidate Kinase: An Enzyme Required for the Biosynthesis of the O-Methyl Phosphoramidate Modification in the Capsular Polysaccharides of Campylobacter jejuni.
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DOI：
10.1021/acs.biochem.8b00279
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发表时间：
2018-04-17
期刊：
影响因子：
2.9
通讯作者：
Raushel FM
中科院分区：
文献类型：
--
作者：
Taylor ZW;Raushel FM
空肠弯曲杆菌是引起胃肠炎的主要原因之一，它产生一种胶囊多糖，该多糖由独特的O-甲基磷酰胺(MeOPN)修饰而成。这种修饰有助于血清抵抗和上皮细胞的侵袭。在此之前，已经阐明了形成MeOPN的前三个生物合成步骤。第一步是由一种新的谷氨酰胺激酶(Cj1418)催化的，它催化依赖三磷酸腺苷的L-谷氨酰胺酰胺氮的磷酸化。L-谷氨酰胺磷酸酶(Cj1416)被谷氨酰胺胞苷转移酶(Cj1416)置换生成胞苷-L-谷氨酰胺，再被Cj1417-谷氨酰胺水解酶(Cj1417)水解生成胞苷二磷酰胺(γ-NH_2)。在这里，我们证明了Cj1415催化依赖于ATP的CDP-NH2的磷酸化生成3‘-磷酸-胞苷-5’-二磷酰胺。Cj1415还将催化二磷酸腺苷(ADP-NH2)和尿苷二磷酰胺(UDP-NH2)的磷酸化，但速度明显降低。建议将Cj1415命名为胞苷二磷酰胺酸激酶。
Campylobacter jejuni , a leading cause of gastroenteritis, produces a capsular polysaccharide that is derivatized with a unique O-methyl phosphoramidate (MeOPN) modification. This modification contributes to serum resistance and invasion of epithelial cells. Previously, the first three biosynthetic steps for the formation of MeOPN have been elucidated. The first step is catalyzed by a novel glutamine kinase (Cj1418), which catalyzes the ATP-dependent phosphorylation of the amide nitrogen of L-glutamine. L-Glutamine phosphate is used by CTP:phosphoglutamine cytidylyltransferase (Cj1416) to displace pyrophosphate from CTP to generate CDP-L-glutamine, which is then hydrolyzed by γ-glutamyl-CDP-amidate hydrolase (Cj1417) to form cytidine diphosphoramidate (CDP-NH2). Here we show that Cj1415 catalyzes the ATP-dependent phosphorylation of CDP-NH2 to form 3′-phospho-cytidine-5′-diphosphoramidate. Cj1415 will also catalyze the phosphorylation of adenosine diphosphoramidate (ADP-NH2) and uridine diphosphoamidate (UDP-NH2) but at significantly reduced rates. It is proposed that Cj1415 be named cytidine diphosphoramidate kinase.
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