异柠檬酸脱氢酶突变型胶质瘤组织提取物中2-羟基戊二酸的（13）C核磁共振检测条件
Conditions for (13)C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas.
作者信息
Pichumani Kumar, Mashimo Tomoyuki, Baek Hyeon-Man, Ratnakar James, Mickey Bruce, DeBerardinis Ralph J, Maher Elizabeth A, Bachoo Robert M, Malloy Craig R, Kovacs Zoltan
机构信息
Advanced Imaging Research Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Annette G. Strauss Center for Neuro-Oncology, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
出版信息
Anal Biochem. 2015 Jul 15;481:4-6. doi: 10.1016/j.ab.2015.04.017. Epub 2015 Apr 20.
(13)C NMR (nuclear magnetic resonance) spectroscopy of extracts from patient tumor samples provides rich information about metabolism. However, in isocitrate dehydrogenase (IDH)-mutant gliomas, (13)C labeling is obscured in oncometabolite 2-hydroxyglutaric acid (2 HG) by glutamate and glutamine, prompting development of a simple method to resolve the metabolites. J-coupled multiplets in 2 HG were similar to glutamate and glutamine and could be clearly resolved at pH 6. A cryogenically cooled (13)C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2 HG. These methods enable the monitoring of (13)C-(13)C spin-spin couplings in 2 HG expressing IDH-mutant gliomas.
对患者肿瘤样本提取物进行的碳-13核磁共振（(13)C NMR）光谱分析可提供有关新陈代谢的丰富信息。然而，在异柠檬酸脱氢酶（IDH）突变型胶质瘤中，谷氨酸和谷氨酰胺会使肿瘤代谢物2-羟基戊二酸（2HG）中的碳-13标记变得模糊不清，这促使人们开发一种简单方法来解析这些代谢物。2HG中的J耦合多重峰与谷氨酸和谷氨酰胺相似，在pH值为6时可以清晰分辨。低温冷却的碳-13探头，而非J分辨异核单量子相干光谱，能显著提高对2HG的检测。这些方法能够监测表达IDH突变型胶质瘤的2HG中的碳-13-碳-13自旋-自旋耦合。
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