Difference between revisions of "Rasmussen 2000 Mol Cell Biochem"
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{{Publication | {{Publication | ||
|title=Rasmussen UF, Rasmussen HN (2000) Human quadriceps muscle mitochondria: a functional characterization. Mol. Cell. Biochem. 208: 37-44. | |title=Rasmussen UF, Rasmussen HN (2000) Human quadriceps muscle mitochondria: a functional characterization. Mol. Cell. Biochem. 208: 37-44. | ||
|info=[http://www.ncbi.nlm.nih.gov/pubmed/10939626 PMID:10939626] | |||
|authors=Rasmussen UF, Rasmussen HN | |authors=Rasmussen UF, Rasmussen HN | ||
|year=2000 | |year=2000 | ||
|journal=Mol. Cell. Biochem. | |journal=Mol. Cell. Biochem. | ||
|abstract=Human quadriceps mitochondria were isolated from ca. 80 mg tissue in ca. 45% yield. The preparation is described with respect to content of mitochondrial markers and nine different respiratory activities. The specific [[ | |abstract=Human quadriceps mitochondria were isolated from ca. 80 mg tissue in ca. 45% yield. The preparation is described with respect to content of mitochondrial markers and nine different respiratory activities. The specific [[State 3]] activities were high in comparison with literature data, indicating high integrity and purity of the preparation. Examples of State 3 rates, in micromol O min(-1) g protein(-1) (25 °C): pyruvate + malate, 400; succinate, 514; malate + glutamate, 444. The notion of high integrity was also supported by the reproducibility of the preparation and the magnitude of the respiratory control ratios and the P/O ratios. The mitochondria most likely had lost ca. 30% of their cytochrome c upon isolation, but it was substantiated that this loss had not influenced the state 3 rates. Functional assays of single reactions or groups of reactions could be based on respiration experiments. The respiratory chain activity, for instance, was measured as respiration of NADH in freeze-permeabilized mitochondria (1263 micromol O min(-1) g protein(-1)). Comparison of uncoupled rates of respiration and state 3 rates indicated that the ATP synthesis exerted major flux control over respiration of succinate + glutamate, malate + glutamate and pyruvate + malate. These reactions, showing very similar rates of ATP synthesis, could be used as a functional assay of ATP synthesis (1200 micromol ATP min(-1) g protein(-1)). Respiration of succinate, palmitoyl-carnitine + malate, or glutamate could not support the maximal rate of ATP synthesis and the upstream reactions probably exerted major flux control in these cases. The specific activities appeared very constant in this group of young men, only the respiratory activity with glutamate might show biological variation. | ||
| | |discipline=Mitochondrial Physiology | ||
}} | }} | ||
{{Labeling | {{Labeling | ||
|organism=Human | |organism=Human | ||
|tissues=Skeletal Muscle | |tissues=Skeletal Muscle | ||
|preparations=Isolated Mitochondria | |preparations=Isolated Mitochondria | ||
|topics=Respiration; OXPHOS; ETS Capacity, Flux Control; Additivity; Threshold; Excess Capacity, Substrate; Glucose; TCA Cycle | |topics=Respiration; OXPHOS; ETS Capacity, Flux Control; Additivity; Threshold; Excess Capacity, Substrate; Glucose; TCA Cycle | ||
|discipline=Mitochondrial Physiology | |||
}} | }} | ||
Revision as of 17:51, 6 November 2011
| Rasmussen UF, Rasmussen HN (2000) Human quadriceps muscle mitochondria: a functional characterization. Mol. Cell. Biochem. 208: 37-44. |
Rasmussen UF, Rasmussen HN (2000) Mol. Cell. Biochem.
Abstract: Human quadriceps mitochondria were isolated from ca. 80 mg tissue in ca. 45% yield. The preparation is described with respect to content of mitochondrial markers and nine different respiratory activities. The specific State 3 activities were high in comparison with literature data, indicating high integrity and purity of the preparation. Examples of State 3 rates, in micromol O min(-1) g protein(-1) (25 °C): pyruvate + malate, 400; succinate, 514; malate + glutamate, 444. The notion of high integrity was also supported by the reproducibility of the preparation and the magnitude of the respiratory control ratios and the P/O ratios. The mitochondria most likely had lost ca. 30% of their cytochrome c upon isolation, but it was substantiated that this loss had not influenced the state 3 rates. Functional assays of single reactions or groups of reactions could be based on respiration experiments. The respiratory chain activity, for instance, was measured as respiration of NADH in freeze-permeabilized mitochondria (1263 micromol O min(-1) g protein(-1)). Comparison of uncoupled rates of respiration and state 3 rates indicated that the ATP synthesis exerted major flux control over respiration of succinate + glutamate, malate + glutamate and pyruvate + malate. These reactions, showing very similar rates of ATP synthesis, could be used as a functional assay of ATP synthesis (1200 micromol ATP min(-1) g protein(-1)). Respiration of succinate, palmitoyl-carnitine + malate, or glutamate could not support the maximal rate of ATP synthesis and the upstream reactions probably exerted major flux control in these cases. The specific activities appeared very constant in this group of young men, only the respiratory activity with glutamate might show biological variation.
Labels:
Organism: Human
Tissue;cell: Skeletal Muscle"Skeletal Muscle" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.
Preparation: Isolated Mitochondria"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.
Regulation: Respiration; OXPHOS; ETS Capacity"Respiration; OXPHOS; ETS Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., Flux Control; Additivity; Threshold; Excess Capacity"Flux Control; Additivity; Threshold; Excess Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property., Substrate; Glucose; TCA Cycle"Substrate; Glucose; TCA Cycle" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
