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. 1988 Mar;81(3):844–852. doi: 10.1172/JCI113393

Fatty acid uptake by isolated rat heart myocytes represents a carrier-mediated transport process.

W Stremmel 1
PMCID: PMC442535  PMID: 3343344

Abstract

The mechanism by which fatty acids enter cardiomyocytes is unclear. Therefore, the influx kinetics of [3H]oleate into isolated rat heart myocytes were examined. Cells were incubated at 37 degrees C with [3H]oleate bound to albumin in various molar ratios and the initial rate of uptake (V0) was determined as a function of the unbound oleate concentration in the medium. V0 was saturable with increasing oleate concentrations incubated (Km 78 nM; Vmax 1.9 nmol X min-1 per 10(6) cells) and temperature dependent with an optimum at 37 degrees C. Furthermore, binding of [3H]oleate to isolated plasma membranes of cardiomyocytes was saturable, revealing a KD of 42 nM, and was inhibited by heat denaturation or trypsin pretreatment of the membranes. From these membranes a single 40-kD protein with high affinity for a variety of long chain fatty acids was isolated. With a monospecific antibody to this membrane protein, binding as well as cellular influx of [3H]oleate was selectively inhibited. These data indicate that at least a portion of myocardial fatty acid uptake is mediated by a specific membrane protein.

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Selected References

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  1. Abumrad N. A., Perkins R. C., Park J. H., Park C. R. Mechanism of long chain fatty acid permeation in the isolated adipocyte. J Biol Chem. 1981 Sep 10;256(17):9183–9191. [PubMed] [Google Scholar]
  2. Altschuld R., Gibb L., Ansel A., Hohl C., Kruger F. A., Brierley G. P. Calcium tolerance of isolated rat heart cells. J Mol Cell Cardiol. 1980 Dec;12(12):1383–1395. doi: 10.1016/0022-2828(80)90123-6. [DOI] [PubMed] [Google Scholar]
  3. Baudhuin P., Evrard P., Berthet J. Electron microscopic examination of subcellular fractions. I. The preparation of representative samples from suspensions of particles. J Cell Biol. 1967 Jan;32(1):181–191. doi: 10.1083/jcb.32.1.181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cleland W. W. Statistical analysis of enzyme kinetic data. Methods Enzymol. 1979;63:103–138. doi: 10.1016/0076-6879(79)63008-2. [DOI] [PubMed] [Google Scholar]
  5. DeGrella R. F., Light R. J. Uptake and metabolism of fatty acids by dispersed adult rat heart myocytes. I. Kinetics of homologous fatty acids. J Biol Chem. 1980 Oct 25;255(20):9731–9738. [PubMed] [Google Scholar]
  6. DeGrella R. F., Light R. J. Uptake and metabolism of fatty acids by dispersed adult rat heart myocytes. II. Inhibition by albumin and fatty acid homologues, and the effect of temperature and metabolic reagents. J Biol Chem. 1980 Oct 25;255(20):9739–9745. [PubMed] [Google Scholar]
  7. Dow J. W., Harding N. G., Powell T. Isolated cardiac myocytes. I. Preparation of adult myocytes and their homology with the intact tissue. Cardiovasc Res. 1981 Sep;15(9):483–514. doi: 10.1093/cvr/15.9.483. [DOI] [PubMed] [Google Scholar]
  8. EVANS J. R., OPIE L. H., SHIPP J. C. METABOLISM OF PALMITIC ACID IN PERFUSED RAT HEART. Am J Physiol. 1963 Oct;205:766–770. doi: 10.1152/ajplegacy.1963.205.4.766. [DOI] [PubMed] [Google Scholar]
  9. Gerards P., Graf W., Kammermeier Glucose transfer studies in isolated cardiocytes of adult rats. J Mol Cell Cardiol. 1982 Mar;14(3):141–149. doi: 10.1016/0022-2828(82)90112-2. [DOI] [PubMed] [Google Scholar]
  10. Glatz J. F., Janssen A. M., Baerwaldt C. C., Veerkamp J. H. Purification and characterization of fatty-acid-binding proteins from rat heart and liver. Biochim Biophys Acta. 1985 Oct 23;837(1):57–66. doi: 10.1016/0005-2760(85)90085-2. [DOI] [PubMed] [Google Scholar]
  11. KOCHWA S., ROSENFIELD R. E., TALLAL L., WASSERMAN L. R. Isoagglutinins associated with ABO erythroblastosis. J Clin Invest. 1961 May;40:874–883. doi: 10.1172/JCI104322. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Langer G. A. Sodium-calcium exchange in the heart. Annu Rev Physiol. 1982;44:435–449. doi: 10.1146/annurev.ph.44.030182.002251. [DOI] [PubMed] [Google Scholar]
  13. Michell R. H., Hawthorne J. N. The site of diphosphoinositide synthesis in rat liver. Biochem Biophys Res Commun. 1965 Nov 22;21(4):333–338. doi: 10.1016/0006-291x(65)90198-1. [DOI] [PubMed] [Google Scholar]
  14. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  15. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed] [Google Scholar]
  16. Paris S., Samuel D., Jacques Y., Gache C., Franchi A., Ailhaud G. The role of serum albumin in the uptake of fatty acids by cultured cardiac cells from chick embryo. Eur J Biochem. 1978 Feb 1;83(1):235–243. doi: 10.1111/j.1432-1033.1978.tb12088.x. [DOI] [PubMed] [Google Scholar]
  17. Piper H. M., Probst I., Schwartz P., Hütter F. J., Spieckermann P. G. Culturing of calcium stable adult cardiac myocytes. J Mol Cell Cardiol. 1982 Jul;14(7):397–412. doi: 10.1016/0022-2828(82)90171-7. [DOI] [PubMed] [Google Scholar]
  18. Powell T., Terrar D. A., Twist V. W. Electrical properties of individual cells isolated from adult rat ventricular myocardium. J Physiol. 1980 May;302:131–153. doi: 10.1113/jphysiol.1980.sp013234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Putney J. W., Jr Stimulus-permeability coupling: role of calcium in the receptor regulation of membrane permeability. Pharmacol Rev. 1978 Jun;30(2):209–245. [PubMed] [Google Scholar]
  20. Said B., Schulz H. Fatty acid binding protein from rat heart. The fatty acid binding proteins from rat heart and liver are different proteins. J Biol Chem. 1984 Jan 25;259(2):1155–1159. [PubMed] [Google Scholar]
  21. Samuel D., Paris S., Ailhaud G. Uptake and metabolism of fatty acids and analogues by cultured cardiac cells from chick embryo. Eur J Biochem. 1976 May 1;64(2):583–595. doi: 10.1111/j.1432-1033.1976.tb10338.x. [DOI] [PubMed] [Google Scholar]
  22. Schwartz A., Allen J. C., Harigaya S. Possible involvement of cardiac Na+, K+-adenosine triphosphatase in the mechanism of action of cardiac glycosides. J Pharmacol Exp Ther. 1969 Jul;168(1):31–41. [PubMed] [Google Scholar]
  23. Skou J. C. Effect of ATP on the intermediary steps of the reaction of the (Na+ plus K+)-dependent enzyme system. 3. Effect on the p-nitrophenylphosphatase activity of the system. Biochim Biophys Acta. 1974 Mar 15;339(2):258–273. [PubMed] [Google Scholar]
  24. Spector A. A., Fletcher J. E., Ashbrook J. D. Analysis of long-chain free fatty acid binding to bovine serum albumin by determination of stepwise equilibrium constants. Biochemistry. 1971 Aug 17;10(17):3229–3232. doi: 10.1021/bi00793a011. [DOI] [PubMed] [Google Scholar]
  25. Stremmel W., Berk P. D. Hepatocellular influx of [14C]oleate reflects membrane transport rather than intracellular metabolism or binding. Proc Natl Acad Sci U S A. 1986 May;83(10):3086–3090. doi: 10.1073/pnas.83.10.3086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stremmel W., Lotz G., Niederau C., Teschke R., Strohmeyer G. Iron uptake by rat duodenal microvillous membrane vesicles: evidence for a carrier mediated transport system. Eur J Clin Invest. 1987 Apr;17(2):136–145. doi: 10.1111/j.1365-2362.1987.tb02393.x. [DOI] [PubMed] [Google Scholar]
  27. Stremmel W., Strohmeyer G., Berk P. D. Hepatocellular uptake of oleate is energy dependent, sodium linked, and inhibited by an antibody to a hepatocyte plasma membrane fatty acid binding protein. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3584–3588. doi: 10.1073/pnas.83.11.3584. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stremmel W., Strohmeyer G., Borchard F., Kochwa S., Berk P. D. Isolation and partial characterization of a fatty acid binding protein in rat liver plasma membranes. Proc Natl Acad Sci U S A. 1985 Jan;82(1):4–8. doi: 10.1073/pnas.82.1.4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stremmel W., Theilmann L. Selective inhibition of long-chain fatty acid uptake in short-term cultured rat hepatocytes by an antibody to the rat liver plasma membrane fatty acid-binding protein. Biochim Biophys Acta. 1986 Jun 11;877(1):191–197. doi: 10.1016/0005-2760(86)90134-7. [DOI] [PubMed] [Google Scholar]
  30. Stremmel W. Translocation of fatty acids across the basolateral rat liver plasma membrane is driven by an active potential-sensitive sodium-dependent transport system. J Biol Chem. 1987 May 5;262(13):6284–6289. [PubMed] [Google Scholar]
  31. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wosilait W. D., Nagy P. A method of computing drug distribution in plasma using stepwise association constants: clofibrate acid as an illustrative example. Comput Programs Biomed. 1976 Oct;6(3):142–148. doi: 10.1016/0010-468x(76)90020-9. [DOI] [PubMed] [Google Scholar]
  33. van Alstyne E., Burch R. M., Knickelbein R. G., Hungerford R. T., Gower E. J., Webb J. G., Poe S. L., Lindenmayer G. E. Isolation of sealed vesicles highly enriched with sarcolemma markers from canine ventricle. Biochim Biophys Acta. 1980 Oct 16;602(1):131–143. doi: 10.1016/0005-2736(80)90296-5. [DOI] [PubMed] [Google Scholar]

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