Mass isotopomer study of anaplerosis from propionate in the perfused rat heart

doi:10.1016/j.abb.2007.02.022

Elsevier

Archives of Biochemistry and Biophysics

Volume 463, Issue 1, 1 July 2007, Pages 110-117

Archives of Biochemistry and Biophysics

https://doi.org/10.1016/j.abb.2007年02月02日2 Get rights and content

Abstract

Anaplerosis from propionate was investigated in rat hearts perfused with 0–2 mM [¹³C₃]propionate and physiological concentrations of glucose, lactate, and pyruvate. The data show that when the concentration of [¹³C₃]propionate was raised from 0 to 2 mM, total anaplerosis increased from 5% to 16% of the turnover of citric acid cycle intermediates. Then, [¹³C₃]propionate abolished anaplerosis from endogenous substrates, glucose, lactate, and pyruvate. Also, while the contents of propionyl-CoA and methylmalonyl-CoA increased with [¹³C₃]propionate concentration, the content of succinyl-CoA decreased, presumably via activation of succinyl-CoA hydrolysis by a decrease in free CoA. Under our conditions, [¹³C₃]propionate was a purely anaplerotic substrate since there was no labeling of mitochondrial acetyl-CoA, reflected by the labeling of the acetyl moiety of citrate.

Section snippets

Materials

Chemicals and biochemicals were obtained from Sigma-Aldrich. [¹³C₃]Propionate, [3-¹³C]propionate, [²H₅]propionate, [¹³C₄]succinic acid, [²H₄]succinic anhydride, [¹³C₄]fumaric acid, and [¹³C₆]citric acid were purchased from Isotec (Miamisburg, OH). [²H₅]Propionyl-CoA and [2,2,3,3-²H₄]succinyl-CoA were prepared by reacting [²H₅]propionic acid and [²H₄]succinic anhydride with CoA [15]. The two labeled acyl-CoAs were purified by HPLC and their water solutions standardized by UV spectroscopy [15].

Results and discussion

This study included two groups of experiments. First, we perfused hearts with 0.25 mM [¹³C₃]propionate for increasing amount of time to follow the time course of metabolite labeling. Second, we perfused hearts with increasing concentrations of [¹³C₃]propionate.

Acknowledgments

This work was supported by grants from the NIH (DK069752), the American Heart Association Ohio Chapter (0465221B) and the Cleveland Mt. Sinai Health Care Foundation.

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Acetate, propionate and butyrate are generated at high levels by colonic bacterial degradation of unabsorbed starch and non-starch polysaccharides and account for a major fraction of energy consumption by epithelial cells (reviewed in Wong et al., 2006; Hijova and Chmelarova, 2007). Several studies have reported utilization of propionate as a favoured energy substrate by heart cells with anaplerotic rates from 0.25 mM propionate estimated to account for 6–8% of the Kreb’s cycle turnover (Kasumov et al., 2007) and other estimates as high as 29% (Sherry et al., 1988). Propionate was shown to be readily utilized as a substrate by Caco-2 colon cancer cells (Malaisse et al., 1996) and was also readily used by the murine mammary carcinoma/sarcoma EMT6 spheroids as an anapleurotic substrate.
Mitochondria are emerging as idealized targets for anti-cancer drugs. One reason for this is that although these organelles are inherent to all cells, drugs are being developed that selectively target the mitochondria of malignant cells without adversely affecting those of normal cells. Such anti-cancer drugs destabilize cancer cell mitochondria and these compounds are referred to as mitocans, classified into several groups according to their mode of action and the location or nature of their specific drug targets. Many mitocans selectively interfere with the bioenergetic functions of cancer cell mitochondria, causing major disruptions often associated with ensuing overloads in ROS production leading to the induction of the intrinsic apoptotic pathway. This in-depth review describes the bases for the bioenergetic differences found between normal and cancer cell mitochondria, focussing on those essential changes occurring during malignancy that clinically may provide the most effective targets for mitocan development. A common theme emerging is that mitochondrially mediated ROS activation as a trigger for apoptosis offers a powerful basis for cancer therapy. Continued research in this area is likely to identify increasing numbers of novel agents that should prove highly effective against a variety of cancers with preferential toxicity towards malignant tissue, circumventing tumor resistance to the other more established therapeutic anti-cancer approaches.
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^☆: Mass isotopomers are designated as M, M1, M2,... Mn, where n is the number of ¹³C or 2H atoms in the ion.

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Archives of Biochemistry and Biophysics

Abstract

Section snippets

Materials

Results and discussion

Acknowledgments

References (37)

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Cited by (22)

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Mass isotopomer study of anaplerosis from propionate in the perfused rat heart☆

Abstract

Section snippets

Materials

Results and discussion

Acknowledgments

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