Protocols and applications of cellular metabolomics in safety studies using precision-cut tissue slices and carbon 13 NMR.
Methods Mol Biol. 2011;691:205-25
Authors: Baverel G, Renault S, Faiz H, El Hage M, Gauthier C, Duplany A, Ferrier B, Martin G
Numerous xenobiotics are toxic to human and animal cells by interacting with their metabolism, but the precise metabolic step affected and the biochemical mechanism behind such a toxicity often remain unknown. In an attempt to reduce the ignorance in this field, we have developed a new approach called cellular metabolomics. This approach, developed in vitro, provides a panoramic view not only of the pathways involved in the metabolism of physiologic substrates of any normal or pathologic human or animal cell but also of the beneficial and adverse effects of xenobiotics on these metabolic pathways. Unlike many cell lines, precision-cut tissue slices, for which there is a renewed interest, remain metabolically differentiated for at least 24-48 h and allow to study the effect of xenobiotics during short-term and long-term incubations. Cellular metabolomics (or cellular metabonomics), which combines enzymatic and carbon 13 NMR measurements with mathematical modeling of metabolic pathways, is illustrated in this brief chapter for studying the effect of insulin on glucose metabolism in rat liver precision-cut slices, and of valproate on glutamine metabolism in human renal cortical precision-cut slices. The use of very small amounts of test compounds allows to predict their toxic effect and eventually their beneficial effects very early in the research and development processes. Cellular metabolomics is complementary to other omics approaches, but, unlike them, provides functional and dynamic pieces of information by measuring enzymatic fluxes.
NMR-based metabolomics of mammalian cell and tissue cultures
NMR-based metabolomics of mammalian cell and tissue cultures
Abstract NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a quantum mechanical total line shape analysis were utilized to quantify 30 metabolites such as amino acids, Krebs cycle intermediates, activated sugars, cofactors, and others in both media and cell extracts. The impact of bioreactor scale and addition of...
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NMR-based metabolomics of mammalian cell and tissue cultures.
NMR-based metabolomics of mammalian cell and tissue cultures.
NMR-based metabolomics of mammalian cell and tissue cultures.
J Biomol NMR. 2011 Mar 4;
Authors: Aranibar N, Borys M, Mackin NA, Ly V, Abu-Absi N, Abu-Absi S, Niemitz M, Schilling B, Li ZJ, Brock B, Russell RJ, Tymiak A, Reily MD
NMR spectroscopy was used to evaluate growth media and the cellular metabolome in two systems of interest to biomedical research. The first of these was a Chinese hamster ovary cell line engineered to express a recombinant protein. Here, NMR spectroscopy and a...
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03-05-2011 01:02 PM
[NMR paper] Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Related Articles Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Biochemistry. 1994 Mar 22;33(11):3304-11
Authors: Aramini JM, Krygsman PH, Vogel HJ
We have examined the binding of Tl3+ to human serotransferrin and chicken ovotransferrin in the presence of carbonate and oxalate by 205Tl and 13C NMR spectroscopy. With carbonate as the synergistic anion, one observes two 205Tl NMR signals due to the bound metal ion in the two...
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08-22-2010 03:33 AM
[NMR paper] Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Related Articles Thallium-205 and carbon-13 NMR studies of human sero- and chicken ovotransferrin.
Biochemistry. 1994 Mar 22;33(11):3304-11
Authors: Aramini JM, Krygsman PH, Vogel HJ
We have examined the binding of Tl3+ to human serotransferrin and chicken ovotransferrin in the presence of carbonate and oxalate by 205Tl and 13C NMR spectroscopy. With carbonate as the synergistic anion, one observes two 205Tl NMR signals due to the bound metal ion in the two...
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08-22-2010 03:33 AM
[NMR paper] NMR studies of fluororetinol analogs complexed to two homologous rat cellular retinol
NMR studies of fluororetinol analogs complexed to two homologous rat cellular retinol-binding proteins.
Related Articles NMR studies of fluororetinol analogs complexed to two homologous rat cellular retinol-binding proteins.
Biochim Biophys Acta. 1994 Sep 21;1208(1):136-44
Authors: Rong D, Lovey AJ, Rosenberger M, d'Avignon DA, Li E
Comparative 19F-NMR studies of fluororetinol analogs with rat cellular retinol binding protein II (CRBP II) and rat cellular retinol-binding protein (CRBP) were performed to probe differences in the binding...
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08-22-2010 03:29 AM
[NMR paper] Carbon-13 NMR studies of the lysine side chains of calmodulin and its proteolytic fra
Carbon-13 NMR studies of the lysine side chains of calmodulin and its proteolytic fragments.
Related Articles Carbon-13 NMR studies of the lysine side chains of calmodulin and its proteolytic fragments.
J Protein Chem. 1993 Dec;12(6):695-707
Authors: Huque ME, Vogel HJ
The pH-titration and dynamic behaviour of the seven lysine side chains in bovine calmodulin were studied by carbon-13 NMR. The amino groups of the calcium saturated protein and its proteolytic fragments TR1C (1-75) and TR2C (78-148) were dimethylated with carbon-13 labeled...
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[NMR paper] Carbon-13 NMR studies of alpha-elastin.
Carbon-13 NMR studies of alpha-elastin.
Related Articles Carbon-13 NMR studies of alpha-elastin.
Ups J Med Sci. 1993;98(1):53-63
Authors: Tarnawski R, Kasperczyk J, Drózdz M
NMR investigations of model protein of elastic fibre is presented. Detailed conformation of alpha-elastin polypeptide chain is discussed by comparison with the conformation of synthetic repeat peptides of elastin. Amino acid composition of alpha-elastin obtained from C-13 NMR spectra correlates with the results of sequencing of tropoelastin.
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[NMR paper] Proton NMR studies on ischemic rat brain tissue.
Proton NMR studies on ischemic rat brain tissue.
Related Articles Proton NMR studies on ischemic rat brain tissue.
Magn Reson Med. 1992 May;25(1):78-84
Authors: Iwama T, Yamada H, Andoh T, Sakai N, Era S, Sogami M, Kuwata K, Watari H
The spin-lattice relaxation time (T1) of water protons and the cross-relaxation time (TIS) between irradiated protein protons and observed water protons were measured in order to study water-macromolecular interactions in ischemic rat brain tissues. Tissues were obtained by bilateral common carotid artery...