Abstract
beta-Hydroxymyristate, -palmitate, and -stearate were produced by and accumulated in isolated rabbit heart when perfused ischemically for 2-10 min by the nonrecirculating langendorff technique with 0.75 mM palmitate and 0.16 mM albumin. Tissue fractionation into mitochondria and cytosol showed that by 2 min of ischemia 44% of beta-hydroxypalmitate and 38% beta-hydroxystearate was located in the cytosol; this percentage increased to greater than 50% by 5 min of ischemia. Lipid fractionation studies showed that by 10 min these two beta-hydroxy fatty acids were distributed approximately as 60% acylcarnitine, 20% acyl-coenzyme A (CoA), and 20% free fatty acids. All three chemical forms of beta-hydroxypalmitate were found in both the mitochondria and the cytosol. After 10 min of ischemia beta-hydroxypalmitoyl-CoA and beta-hydroxystearoyl-CoA constituted at least 16% of the incremental long-chain acyl-CoA, whereas beta-hydroxypalmitoylcarnitine and b-hydroxystearoylcarnitine constituted 8% of the incremental long-chain acylcarnitine. These data suggests that myocardial beta-hydroxyacyl-CoA oxidation is limited during ischemia. Substrate accumulates and is transferred to the cytosol where it accumulates primarily as beta-hydroxyacylcarnitine.
Full text
PDF![377](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/58a78f5ec513/jcinvest00478-0129.png)
![378](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/e850759b70ba/jcinvest00478-0130.png)
![379](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/21521dfdc036/jcinvest00478-0131.png)
![380](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/8b8ea5cd26dd/jcinvest00478-0132.png)
![381](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/320854552643/jcinvest00478-0133.png)
![382](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/85053ffdff5b/jcinvest00478-0134.png)
![383](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5283/370987/c4e5cef502c1/jcinvest00478-0135.png)
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allred J. B., Guy D. G. Determination of coenzyme A and acetyl CoA in tissue extracts. Anal Biochem. 1969 May;29(2):293–299. doi: 10.1016/0003-2697(69)90312-1. [DOI] [PubMed] [Google Scholar]
- BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
- Bremer J., Wojtczak A. B. Factors controlling the rate of fatty acid -oxidation in rat liver mitochondria. Biochim Biophys Acta. 1972 Dec 8;280(4):515–530. doi: 10.1016/0005-2760(72)90131-2. [DOI] [PubMed] [Google Scholar]
- Crass M. F., 3rd, McCaskill E. S., Shipp J. C. Effect of pressure development on glucose and palmitate metabolism in perfused heart. Am J Physiol. 1969 Jun;216(6):1569–1576. doi: 10.1152/ajplegacy.1969.216.6.1569. [DOI] [PubMed] [Google Scholar]
- Crass M. F., 3rd, Pieper G. M. Lipid and glycogen metabolism in the hypoxic heart: effects of epinephrine. Am J Physiol. 1975 Oct;229(4):885–889. doi: 10.1152/ajplegacy.1975.229.4.885. [DOI] [PubMed] [Google Scholar]
- Hull F. E., Radloff J. F., Sweeley C. C. beta-Hydroxy fatty acid production during fatty acid oxidation by heart mitochondria. Recent Adv Stud Cardiac Struct Metab. 1975;7:13–21. [PubMed] [Google Scholar]
- Idell-Wenger J. A., Grotyohann L. W., Neely J. R. Coenzyme A and carnitine distribution in normal and ischemic hearts. J Biol Chem. 1978 Jun 25;253(12):4310–4318. [PubMed] [Google Scholar]
- Kako K. J., Patterson S. D. Phosphatidate phosphohydrolase and palmitoyl-coenzyme A hydrolase in cardiac subcellular fractions of hyperthyroid rabbits and cardiomyopathic hamsters. Biochem J. 1975 Nov;152(2):313–323. doi: 10.1042/bj1520313. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kurooka S., Hosoki K., Yoshimura Y. Some properties of long fatty acyl-coenzyme A thioesterase in rat organs. J Biochem. 1972 Apr;71(4):625–634. [PubMed] [Google Scholar]
- Lochner A., Kotzé J. C., Benade A. J., Gevers W. Mitochondrial oxidative phosphorylation in low-flow hypoxia: role of free fatty acids. J Mol Cell Cardiol. 1978 Sep;10(9):857–875. doi: 10.1016/0022-2828(78)90394-2. [DOI] [PubMed] [Google Scholar]
- Lopes-Cardozo M., Klazinga W., van den Bergh S. G. Accumulation of carnitine esters of beta-oxidation intermediates during palmitate oxidation by rat-liver mitochondria. Eur J Biochem. 1978 Feb;83(2):629–634. doi: 10.1111/j.1432-1033.1978.tb12132.x. [DOI] [PubMed] [Google Scholar]
- Mancha M., Stokes G. B., Stumpf P. K. Fat metabolism in higher plants. The determination of acyl-acyl carrier protein and acyl coenzyme A in a complex lipid mixture 1,2. Anal Biochem. 1975 Oct;68(2):600–608. doi: 10.1016/0003-2697(75)90655-7. [DOI] [PubMed] [Google Scholar]
- Matlib M. A., Rebman D., Ashraf M., Rouslin W., Schwartz A. Differential activities of putative subsarcolemmal and interfibrillar mitochondria from cardiac muscle. J Mol Cell Cardiol. 1981 Feb;13(2):163–170. doi: 10.1016/0022-2828(81)90213-3. [DOI] [PubMed] [Google Scholar]
- McGarry J. D., Foster D. W. An improved and simplified radioisotopic assay for the determination of free and esterified carnitine. J Lipid Res. 1976 May;17(3):277–281. [PubMed] [Google Scholar]
- Moore K. H., Radloff J. F., Hull F. E., Sweeley C. C. Incomplete fatty acid oxidation by ischemic heart: beta-hydroxy fatty acid production. Am J Physiol. 1980 Aug;239(2):H257–H265. doi: 10.1152/ajpheart.1980.239.2.H257. [DOI] [PubMed] [Google Scholar]
- Neely J. R., Rovetto M. J., Whitmer J. T. Rate-limiting steps of carbohydrate and fatty acid metabolism in ischemic hearts. Acta Med Scand Suppl. 1976;587:9–15. doi: 10.1111/j.0954-6820.1976.tb05861.x. [DOI] [PubMed] [Google Scholar]
- Oram J. F., Bennetch S. L., Neely J. R. Regulation of fatty acid utilization in isolated perfused rat hearts. J Biol Chem. 1973 Aug 10;248(15):5299–5309. [PubMed] [Google Scholar]
- Palmer J. W., Tandler B., Hoppel C. L. Biochemical properties of subsarcolemmal and interfibrillar mitochondria isolated from rat cardiac muscle. J Biol Chem. 1977 Dec 10;252(23):8731–8739. [PubMed] [Google Scholar]
- Pande S. V. On rate-controlling factors of long chain fatty acid oxidation. J Biol Chem. 1971 Sep 10;246(17):5384–5390. [PubMed] [Google Scholar]
- Shug A. L., Shrago E., Bittar N., Folts J. D., Koke J. R. Acyl-CoA inhibition of adenine nucleotide translocation in ischemic myocardium. Am J Physiol. 1975 Mar;228(3):689–692. doi: 10.1152/ajplegacy.1975.228.3.689. [DOI] [PubMed] [Google Scholar]
- Sobel B. E., Corr P. B., Robison A. K., Goldstein R. A., Witkowski F. X., Klein M. S. Accumulation of lysophosphoglycerides with arrhythmogenic properties in ischemic myocardium. J Clin Invest. 1978 Sep;62(3):546–553. doi: 10.1172/JCI109159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sordahl L. A., Schwartz A. Effects of dipyridamole on heart muscle mitochondria. Mol Pharmacol. 1967 Nov;3(6):509–515. [PubMed] [Google Scholar]
- Stanley K. K., Tubbs P. K. The role of intermediates in mitochondrial fatty acid oxidation. Biochem J. 1975 Jul;150(1):77–88. doi: 10.1042/bj1500077. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Veloso D., Veech R. L. Stoichiometric hydrolysis of long chain acyl-CoA and measurement of the CoA formed with an enzymatic cycling method. Anal Biochem. 1974 Dec;62(2):449–450. doi: 10.1016/0003-2697(74)90177-8. [DOI] [PubMed] [Google Scholar]
- Whitmer J. T., Idell-Wenger J. A., Rovetto M. J., Neely J. R. Control of fatty acid metabolism in ischemic and hypoxic hearts. J Biol Chem. 1978 Jun 25;253(12):4305–4309. [PubMed] [Google Scholar]
- Wood J. M., Bush B., Pitts B. J., Schwartz A. Inhibition of bovine heart Na+, K+-ATPase by palmitylcarnitine and palmityl-CoA. Biochem Biophys Res Commun. 1977 Jan 24;74(2):677–684. doi: 10.1016/0006-291x(77)90356-4. [DOI] [PubMed] [Google Scholar]
- Wood J. M., Wallick E. T., Schwartz A., Chang C. H. The effect of palmitoyl-coenzyme A on rat heart and liver mitochondria. Oxygen consumption and palmitoylcarnitine formation. Biochim Biophys Acta. 1977 Feb 23;486(2):331–340. doi: 10.1016/0005-2760(77)90029-7. [DOI] [PubMed] [Google Scholar]