Abstract Mitochondrial toxicity has been a serious concern, not only in preclinical drug development but also in clinical trials. In mitochondria, there are several distinct metabolic processes including fatty acid β-oxidation, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS), and each process contains discrete but often intimately linked steps. Interruption in any one of those steps can cause mitochondrial dysfunction. Detection of inhibition to OXPHOS can be complicated in vivo because intermediate endogenous metabolites can be recycled in situ or circulated systemically for metabolism in other organs or tissues. Commonly used assays for evaluating mitochondrial function are often applied to ex vivo or in vitro samples; they include various enzymatic or protein assays, as well as functional assays such as measurement of oxygen consumption rate, membrane potential, or acidification rates. Metabolomics provides quantitative profiles of overall metabolic changes that can aid in the unraveling of explicit biochemical details of mitochondrial inhibition while providing a holistic view and heuristic understanding of cellular bioenergetics. In this paper, we showed the application of quantitative NMR metabolomics to in vitro myotube cells treated with mitochondrial toxicants, rotenone and antimycin A. The close coupling of the TCA cycle to the electron transfer chain (ETC) in OXPHOS enables specific diagnoses of inhibition to ETC complexes by discrete biochemical changes in the TCA cycle.
Content Type Journal Article
Pages 1-13
DOI 10.1007/s10858-011-9482-8
Authors
Qiuwei Xu, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
Heather Vu, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
Liping Liu, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
Ting-Chuan Wang, Merck Research Laboratories, 126 E. Lincoln Avenue, Rahway, NJ 07065, USA
William H. Schaefer, Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA
Comparative analysis of essential collective dynamics and NMR-derived flexibility profiles in evolutionarily diverse prion proteins.
Comparative analysis of essential collective dynamics and NMR-derived flexibility profiles in evolutionarily diverse prion proteins.
Comparative analysis of essential collective dynamics and NMR-derived flexibility profiles in evolutionarily diverse prion proteins.
Prion. 2011 Jul 1;5(3)
Authors: Santo KP, Berjanskii M, Wishart DS, Stepanova M
Abstract
Collective motions on ns-?s time scales are known to have a major impact on protein folding, stability, binding and enzymatic efficiency. It is also believed that these motions may have an...
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08-27-2011 04:53 PM
Mutations in the Saccharomyces cerevisiae succinate dehydrogenase result in distinct metabolic phenotypes revealed through (1)H NMR-based metabolic footprinting.
Mutations in the Saccharomyces cerevisiae succinate dehydrogenase result in distinct metabolic phenotypes revealed through (1)H NMR-based metabolic footprinting.
Mutations in the Saccharomyces cerevisiae succinate dehydrogenase result in distinct metabolic phenotypes revealed through (1)H NMR-based metabolic footprinting.
J Proteome Res. 2010 Dec 3;9(12):6729-39
Authors: Szeto SS, Reinke SN, Sykes BD, Lemire BD
Metabolomics is a powerful method of examining the intricate connections between mutations, metabolism, and disease. Metabolic...
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05-25-2011 07:01 PM
Changes in the NMR metabolic profile of human microglial cells exposed to lipopolysaccharide or morphine.
Changes in the NMR metabolic profile of human microglial cells exposed to lipopolysaccharide or morphine.
Changes in the NMR metabolic profile of human microglial cells exposed to lipopolysaccharide or morphine.
J Neuroimmune Pharmacol. 2010 Dec;5(4):574-81
Authors: El Ghazi I, Sheng WS, Hu S, Reilly BG, Lokensgard JR, Rock RB, Peterson PK, Wilcox GL, Armitage IM
Microglial cells play a major role in host defense of the central nervous system. Once activated, several functional properties are up-regulated including migration, phagocytosis, and...
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03-01-2011 12:14 PM
[U. of Ottawa NMR Facility Blog] Excitation Profiles for Shaped Pulses
Excitation Profiles for Shaped Pulses
Shaped pulses are very commonly used for selective excitation and nonselective inversion in a large number of NMR pulse sequences. The frequency domain excitation profile of a radio frequency pulse is the Fourier transform of the time dependent pulse shape and determines the width, uniformity and phase of the frequency spectrum excited. Since time and frequency are reciprocals of one another, short rf pulses have very wide excitation profiles and long rf pulses have very narrow selective excitation profiles. In a previous BLOG post the excitation...
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01-21-2011 03:31 AM
[NMR paper] Specific in vitro labeling of cells with a fluorine-19 probe encapsulated in antibody
Specific in vitro labeling of cells with a fluorine-19 probe encapsulated in antibody-targeted liposomes: a F-19 NMR spectroscopy study.
Related Articles Specific in vitro labeling of cells with a fluorine-19 probe encapsulated in antibody-targeted liposomes: a F-19 NMR spectroscopy study.
Magn Reson Med. 1993 Feb;29(2):252-5
Authors: Vion-Dury J, Machy P, Confort-Gouny S, Leserman L, Cozzone PJ
Liposomes containing dexamethasone phosphate (DMp) were covalently coupled to protein A and then incubated with murine L929 fibroblast and RDM4...
Metabolic profiles show specific mitochondrial toxicities in vitro in myotube cells
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