Wisniewski 1995 Eur J Biochem: Difference between revisions
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|year=1995 | |year=1995 | ||
|journal=Eur J Biochem | |journal=Eur J Biochem | ||
|abstract=Metabolic control analysis was applied to describe the control of mitochondrial oxidative phosphorylation | |abstract=Metabolic control analysis was applied to describe the control of mitochondrial oxidative phosphorylation in calcium (approximately 2 pM free calcium) activated saponin-skinned rat musculus soleus fibers oxidizing glutamate and malate. Under these circumstances approximately 80 % of mitochondrial activestate respiration was reached due to the activation of ATP turnover by actomyosin ATPase. The flux control coefficients of H+-ATPase, adenine-nucleotide translocase, phosphate transporter, NADH :ubiquinone oxidoreductase and cytochrome-c oxidase were determined to be equal to 0.16 2 0.08 (n = 6), 0.34? 0.12 (n = 5), 0.08 2 0.03 (n = 5), 0.01 -+ 0.006 (n = 4) and 0.09 ? 0.03 (n = 3) using inhibitor titrations with the specific inhibitors oligomycin, carboxyatractyloside, mersalyl, rotenone and cyanide, respectively, and applying non-linear regression of the entire titration curve. The flux control coefficient of actomyosin ATPase was determined with vanadate to be equal to 0.50?0.09 (n = 6), measuring independently the vanadate-caused inhibition of fiber respiration and ATP-splitting activity. In contrast to results with isolated rat skeletal muscle mitochondria reconstituted with soluble F,-ATPase the decrease in phosphate concentration from 10 mM to 1 mM only slightly affected the distribution of flux control coefficients. This difference is caused by different kinetic properties of soluble F,-ATPase and actomyosin ATPase. Therefore, phosphate seems to be in skeletal muscle in vivo only a modest modulator of control of oxidative phosphorylation. | ||
in calcium (approximately 2 pM free calcium) activated saponin-skinned rat musculus soleus fibers | |||
oxidizing glutamate and malate. Under these circumstances approximately 80 % of mitochondrial activestate | |||
respiration was reached due to the activation of ATP turnover by actomyosin ATPase. The flux | |||
control coefficients of H+-ATPase, adenine-nucleotide translocase, phosphate transporter, NADH :ubiquinone | |||
oxidoreductase and cytochrome-c oxidase were determined to be equal to 0.16 2 0.08 (n = 6), | |||
0.34? 0.12 (n = 5), 0.08 2 0.03 (n = 5), 0.01 -+ 0.006 (n = 4) and 0.09 ? 0.03 (n = 3) using inhibitor | |||
titrations with the specific inhibitors oligomycin, carboxyatractyloside, mersalyl, rotenone and cyanide, | |||
respectively, and applying non-linear regression of the entire titration curve. The flux control coefficient | |||
of actomyosin ATPase was determined with vanadate to be equal to 0.50?0.09 (n = 6), measuring | |||
independently the vanadate-caused inhibition of fiber respiration and ATP-splitting activity. In contrast to | |||
results with isolated rat skeletal muscle mitochondria reconstituted with soluble F,-ATPase the decrease | |||
in phosphate concentration from 10 mM to 1 mM only slightly affected the distribution of flux control | |||
coefficients. This difference is caused by different kinetic properties of soluble F,-ATPase and actomyosin | |||
ATPase. Therefore, phosphate seems to be in skeletal muscle in vivo only a modest modulator of control | |||
of oxidative phosphorylation. | |||
|keywords=Skinned muscle fibers, Flux control, Oxidative phosphorylation | |keywords=Skinned muscle fibers, Flux control, Oxidative phosphorylation | ||
|mipnetlab=DE Magdeburg Gellerich FN | |||
}} | }} | ||
{{Labeling | {{Labeling | ||
|instruments=Oxygraph-2k | |instruments=Oxygraph-2k | ||
|organism=Rat | |organism=Rat | ||
|tissues=Skeletal | |tissues=Skeletal muscle | ||
|preparations=Permeabilized | |preparations=Permeabilized tissue | ||
}} | }} |
Revision as of 02:57, 5 April 2012
Wisniewski E, Gellerich FN, Kunz WS (1995) Distribution of flux control among the enzymes of mitochondrial oxidative phosphorylation in calcium-activated saponin-skinned rat musculus soleus fibers. Eur J Biochem 230: 549-554. |
Wisniewski E, Gellerich FN, Kunz WS (1995) Eur J Biochem
Abstract: Metabolic control analysis was applied to describe the control of mitochondrial oxidative phosphorylation in calcium (approximately 2 pM free calcium) activated saponin-skinned rat musculus soleus fibers oxidizing glutamate and malate. Under these circumstances approximately 80 % of mitochondrial activestate respiration was reached due to the activation of ATP turnover by actomyosin ATPase. The flux control coefficients of H+-ATPase, adenine-nucleotide translocase, phosphate transporter, NADH :ubiquinone oxidoreductase and cytochrome-c oxidase were determined to be equal to 0.16 2 0.08 (n = 6), 0.34? 0.12 (n = 5), 0.08 2 0.03 (n = 5), 0.01 -+ 0.006 (n = 4) and 0.09 ? 0.03 (n = 3) using inhibitor titrations with the specific inhibitors oligomycin, carboxyatractyloside, mersalyl, rotenone and cyanide, respectively, and applying non-linear regression of the entire titration curve. The flux control coefficient of actomyosin ATPase was determined with vanadate to be equal to 0.50?0.09 (n = 6), measuring independently the vanadate-caused inhibition of fiber respiration and ATP-splitting activity. In contrast to results with isolated rat skeletal muscle mitochondria reconstituted with soluble F,-ATPase the decrease in phosphate concentration from 10 mM to 1 mM only slightly affected the distribution of flux control coefficients. This difference is caused by different kinetic properties of soluble F,-ATPase and actomyosin ATPase. Therefore, phosphate seems to be in skeletal muscle in vivo only a modest modulator of control of oxidative phosphorylation. โข Keywords: Skinned muscle fibers, Flux control, Oxidative phosphorylation
โข O2k-Network Lab: DE Magdeburg Gellerich FN
Labels:
Organism: Rat
Tissue;cell: Skeletal muscle
Preparation: Permeabilized tissue
HRR: Oxygraph-2k