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Drug Discovery Assay to Identify Modulators of the Mitochondrial Ca Uniporter.
Methods Mol Biol
July 2021
Institute for Diabetes and Obesity, Helmholtz Diabetes Center (HDC), Helmholtz Zentrum München and German National Diabetes Center (DZD), Neuherberg, Germany.
The mitochondrial calcium uniporter (MCU ) is an essential protein of the inner mitochondrial membrane that mediates the uptake of calcium into mitochondria of virtually all mammalian tissues, regulating cell metabolism, signaling, and death. MCU-mediated calcium uptake has been shown to play a pathophysiological role in diverse human disease contexts, which qualifies this channel as a druggable target for therapeutic intervention.Here, we present a protocol to perform drug screens to identify effective and specific MCU-targeting inhibitors.
View Article and Find Full Text PDFLactation performance of dairy cows fed rehydrated and ensiled corn grain differing in particle size and proportion in the diet.
J Dairy Sci
November 2019
Better Nature Research Center, Ijaci, MG 37218-000, Brazil; Empresa de Pesquisa Agropecuária de Minas Gerais, Lavras, MG 37200-000, Brazil.
Rehydrated and ensiled mature ground corn has high ruminal starch digestibility, but particle size (PS) and dietary starch proportion (ST) can affect starch digestion and lactating cow performance. We evaluated the effect of rehydrated and ensiled corn (REC), PS, and ST on intake, lactation performance, nutrient digestibility, ruminal fermentation profile, and chewing behavior of dairy cows. Kernels from an 84% vitreousness hybrid were finely (FN) or coarsely (CS) ground, yielding geometric mean particle sizes of 1,591 and 2,185 µm, respectively.
View Article and Find Full Text PDFSystematic Identification of MCU Modulators by Orthogonal Interspecies Chemical Screening.
Mol Cell
August 2017
Gene Center/Department of Biochemistry, Ludwig-Maximilians Universität München, Munich 81377, Germany; Institute for Diabetes and Obesity, Helmholtz Zentrum München, Neuherberg 85764, Germany. Electronic address:
The mitochondrial calcium uniporter complex is essential for calcium (Ca) uptake into mitochondria of all mammalian tissues, where it regulates bioenergetics, cell death, and Ca signal transduction. Despite its involvement in several human diseases, we currently lack pharmacological agents for targeting uniporter activity. Here we introduce a high-throughput assay that selects for human MCU-specific small-molecule modulators in primary drug screens.
View Article and Find Full Text PDFNi2+-activated glyoxalase I from Escherichia coli: substrate specificity, kinetic isotope effects and evolution within the βαβββ superfamily.
J Inorg Biochem
March 2012
Department of Chemistry, University of Waterloo, Waterloo, Ontario, Canada.
The Escherichia coli glyoxalase system consists of the metalloenzymes glyoxalase I and glyoxalase II. Little is known regarding Ni(2+)-activated E. coli glyoxalase I substrate specificity, its thiol cofactor preference, the presence or absence of any substrate kinetic isotope effects on the enzyme mechanism, or whether glyoxalase I might catalyze additional reactions similar to those exhibited by related βαβββ structural superfamily members.
View Article and Find Full Text PDFPurification of electron-transferring flavoprotein from Megasphaera elsdenii and binding of additional FAD with an unusual absorption spectrum.
J Biochem
November 2003
Department of Molecular Physiology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, Kumamoto, 860-8556.
Electron-transferring flavoprotein (ETF), its redox partner flavoproteins, i.e., D-lactate dehydrogenase and butyryl-CoA dehydrogenase, and another well-known flavoprotein, flavodoxin, were purified from the same starting cell paste of an anaerobic bacterium, Megasphaera elsdenii.
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