Conditions for 13C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas

Kumar Pichumani; Tomoyuki Mashimo; Hyeon Man Baek; James Ratnakar; Bruce Mickey; Ralph J. DeBerardinis; Elizabeth A. Maher; Robert M. Bachoo; Craig R. Malloy; Zoltan Kovacs

doi:10.1016/j.ab.2015.04.017

Conditions for ¹³C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas

Kumar Pichumani, Tomoyuki Mashimo, Hyeon Man Baek, James Ratnakar, Bruce Mickey, Ralph J. DeBerardinis, Elizabeth A. Maher, Robert M. Bachoo, Craig R. Malloy, Zoltan Kovacs

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

Abstract

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 cooled ¹³C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of ¹³C-¹³C spin-spin couplings in 2HG expressing IDH-mutant gliomas.

Original language	English (US)
Pages (from-to)	4-6
Number of pages	3
Journal	Analytical Biochemistry
Volume	481
DOIs	https://doi.org/10.1016/j.ab.2015.04.017
State	Published - Jun 3 2015

Keywords

2-Hydroxyglutarate
C NMR
Cryoprobe
IDH-mutant gliomas

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.ab.2015.04.017

Cite this

@article{dabaac5b8a32425fb375be8c9b7a4b24,

title = "Conditions for 13C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas",

abstract = "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 cooled 13C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of 13C-13C spin-spin couplings in 2HG expressing IDH-mutant gliomas.",

keywords = "2-Hydroxyglutarate, C NMR, Cryoprobe, IDH-mutant gliomas",

author = "Kumar Pichumani and Tomoyuki Mashimo and Baek, {Hyeon Man} and James Ratnakar and Bruce Mickey and DeBerardinis, {Ralph J.} and Maher, {Elizabeth A.} and Bachoo, {Robert M.} and Malloy, {Craig R.} and Zoltan Kovacs",

note = "Funding Information: This study was supported by grants from the National Institutes of Health ( P41EB015908 , R01CA154843 ) and the Cancer Prevention Research Institute of Texas ( RP140021-P2 , RP101243 and RP130362 ). Publisher Copyright: {\textcopyright} 2015 Elsevier Inc. All rights reserved.",

year = "2015",

month = jun,

day = "3",

doi = "10.1016/j.ab.2015.04.017",

language = "English (US)",

volume = "481",

pages = "4--6",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press",

}

TY - JOUR

T1 - Conditions for 13C NMR detection of 2-hydroxyglutarate in tissue extracts from isocitrate dehydrogenase-mutated gliomas

AU - Pichumani, Kumar

AU - Mashimo, Tomoyuki

AU - Baek, Hyeon Man

AU - Ratnakar, James

AU - Mickey, Bruce

AU - DeBerardinis, Ralph J.

AU - Maher, Elizabeth A.

AU - Bachoo, Robert M.

AU - Malloy, Craig R.

AU - Kovacs, Zoltan

N1 - Funding Information: This study was supported by grants from the National Institutes of Health ( P41EB015908 , R01CA154843 ) and the Cancer Prevention Research Institute of Texas ( RP140021-P2 , RP101243 and RP130362 ). Publisher Copyright: © 2015 Elsevier Inc. All rights reserved.

PY - 2015/6/3

Y1 - 2015/6/3

N2 - 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 cooled 13C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of 13C-13C spin-spin couplings in 2HG expressing IDH-mutant gliomas.

AB - 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 cooled 13C probe, but not J-resolved heteronuclear single quantum coherence spectroscopy, significantly improved detection of 2HG. These methods enable the monitoring of 13C-13C spin-spin couplings in 2HG expressing IDH-mutant gliomas.

KW - 2-Hydroxyglutarate

KW - C NMR

KW - Cryoprobe

KW - IDH-mutant gliomas

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