Conditions for (13)C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas.
Conditions for (13)C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas.
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(13)C NMR 检测异柠檬酸脱氢酶突变神经胶质瘤组织提取物中 2-羟基戊二酸的条件。
DOI：
10.1016/j.ab.2015.04.017
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发表时间：
2015
影响因子：
2.9
通讯作者：
Kovacs,Zoltan
中科院分区：
文献类型：
--
作者：
Pichumani,Kumar;Mashimo,Tomoyuki;Baek,Hyeon-Man;Ratnakar,James;Mickey,Bruce;DeBerardinis,RalphJ;Maher,ElizabethA;Bachoo,RobertM;Malloy,CraigR;Kovacs,Zoltan
来自患者肿瘤样品的提取物的13 C NMR（核磁共振）光谱提供了关于代谢的丰富信息。然而，在异柠檬酸脱氢酶（IDH）突变型胶质瘤中，13 C标记在癌代谢物2-羟基谷氨酸（2 HG）中被谷氨酸和谷氨酰胺遮蔽，促使开发了一种简单的方法来解析代谢物。在2 HG中的J-偶联多重峰类似于谷氨酸和谷氨酰胺，并且可以在pH 6下清楚地分辨。低温冷却的13 C探针，而不是J-分辨异质结单量子相干光谱，显着提高检测2 HG。这些方法能够监测表达2 HG的IDH突变胶质瘤中的13 C-13 C自旋-自旋耦合。
13C NMR (nuclear magnetic resonance) spectroscopy of extracts from patient tumor samples provides rich information about metabolism. However, in isocitrate dehydrogenase (IDH)-mutant gliomas,13C labeling is obscured in oncometabolite 2-hydroxyglutaric acid (2HG) by glutamate and glutamine, prompting development of a simple method to resolve the metabolites. J-coupled multiplets in 2HG were similar to glutamate and glutamine and could be clearly resolved at pH 6. A cryogenically cooled13C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of13C–13C spin–spin couplings in 2HG expressing IDH-mutant gliomas.