Open Access
Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism
TLDR
In this article, a method for the rapid and specific isolation of mitochondria and use it in tandem with a database of predicted mitochondrial metabolites to measure the matrix concentrations of more than 100 metabolites across various states of respiratory chain (RC) function.Abstract:
Mitochondria house metabolic pathways that impact most aspects of cellular physiology. While metabolite profiling by mass spectrometry is widely applied at the whole-cell level, it is not routinely possible to measure the concentrations of small molecules in mammalian organelles. We describe a method for the rapid and specific isolation of mitochondria and use it in tandem with a database of predicted mitochondrial metabolites ("MITObolome") to measure the matrix concentrations of more than 100 metabolites across various states of respiratory chain (RC) function. Disruption of the RC reveals extensive compartmentalization of mitochondrial metabolism and signatures unique to the inhibition of each RC complex. Pyruvate enables the proliferation of RC-deficient cells but has surprisingly limited effects on matrix contents. Interestingly, despite failing to restore matrix NADH/NAD balance, pyruvate does increase aspartate, likely through the exchange of matrix glutamate for cytosolic aspartate. We demonstrate the value of mitochondrial metabolite profiling and describe a strategy applicable to other organelles.read more
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Methods in Enzymology.
TL;DR: This volume is keyed to high resolution electron microscopy, which is a sophisticated form of structural analysis, but really morphology in a modern guise, the physical and mechanical background of the instrument and its ancillary tools are simply and well presented.
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From Sugar To Acetate - The Origins Of Acetyl-Coa Dictate Its Use In Cells And In Mice
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References
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Journal Article
Methods in Enzymology.
TL;DR: This volume is keyed to high resolution electron microscopy, which is a sophisticated form of structural analysis, but really morphology in a modern guise, the physical and mechanical background of the instrument and its ancillary tools are simply and well presented.
Journal ArticleDOI
Java Treeview---extensible visualization of microarray data
TL;DR: Java Treeview as mentioned in this paper is an open-source, cross-platform rewrite that handles very large datasets well, and supports extensions to the file format that allow the results of additional analysis to be visualized and compared.
Journal ArticleDOI
A Mitochondrial Protein Compendium Elucidates Complex I Disease Biology
David J. Pagliarini,Sarah E. Calvo,Betty Chang,Sunil A Sheth,Scott Bradley Vafai,Shao En Ong,Geoffrey A. Walford,Canny Sugiana,Avihu Boneh,Avihu Boneh,William K. Chen,David E. Hill,Marc Vidal,James G. Evans,David R. Thorburn,David R. Thorburn,Steven A. Carr,Vamsi K. Mootha,Vamsi K. Mootha +18 more
TL;DR: This work predicts 19 proteins to be important for the function of complex I (CI) of the electron transport chain and validate a subset of these predictions using RNAi, including C8orf38, which is shown to have an inherited mutation in a lethal, infantile CI deficiency.
Journal ArticleDOI
Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation
Michael P. King,Giuseppe Attardi +1 more
TL;DR: Transformants obtained with various mitochondrial donors exhibited a respiratory phenotype that was in most cases distinct from that of the rho 0 parent or the donor, indicating that the genotypes of the mitochondrial and nuclear genomes as well as their specific interactions play a role in the respiratory competence of a cell.
Journal ArticleDOI
Reductive carboxylation supports growth in tumour cells with defective mitochondria
Andrew R. Mullen,William W. Wheaton,Eunsook S. Jin,Pei Hsuan Chen,Lucas B. Sullivan,Tzuling Cheng,Youfeng Yang,W. Marston Linehan,Navdeep S. Chandel,Ralph J. DeBerardinis +9 more
TL;DR: These findings reveal the novel induction of a versatile glutamine-dependent pathway that reverses many of the reactions of the canonical CAC, supports tumour cell growth, and explains how cells generate pools of CAC intermediates in the face of impaired mitochondrial metabolism.
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