Legerton T L, Kanamori K, Weiss R L, Roberts J D
Proc Natl Acad Sci U S A. 1981 Mar;78(3):1495-8. doi: 10.1073/pnas.78.3.1495.
Suspensions of intact mycelia of Neurospora crassa grown in medium containing 15NH4Cl have been found to give well-resolved 15N nuclear magnetic resonance spectra for metabolites that play crucial roles in intermediary nitrogen metabolism. These include the amide nitrogen of glutamine, the alpha-amino nitrogens of glutamate and other amino acids, the guanidino nitrogens of arginine, the ureido nitrogen of citrulline, the side-chain nitrogens of ornithine or lysine, or both, and uridine diphosphates. The turnover time of glutamine in vivo was estimated to be less than 1 hr by using nuclear magnetic resonance spectroscopy in conjunction with tracer methodologies. Applications of these techniques to the study of nitrogen metabolism are discussed.
已发现,在含有15NH4Cl的培养基中生长的粗糙脉孢菌完整菌丝体悬浮液,对于在中间氮代谢中起关键作用的代谢物,能给出分辨率良好的15N核磁共振谱。这些代谢物包括谷氨酰胺的酰胺氮、谷氨酸和其他氨基酸的α-氨基氮、精氨酸的胍基氮、瓜氨酸的脲基氮、鸟氨酸或赖氨酸的侧链氮(或两者)以及尿苷二磷酸。通过结合示踪方法使用核磁共振光谱,体内谷氨酰胺的周转时间估计不到1小时。讨论了这些技术在氮代谢研究中的应用。