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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Jan;82(1):4–8. doi: 10.1073/pnas.82.1.4

Isolation and partial characterization of a fatty acid binding protein in rat liver plasma membranes.

W Stremmel, G Strohmeyer, F Borchard, S Kochwa, P D Berk
PMCID: PMC396959  PMID: 3881757

Abstract

When [14C]oleate-bovine serum albumin complexes were incubated in vitro with rat liver plasma membranes (LPM), specific, saturable binding of oleate to the membranes was observed. Maximal heat-sensitive (i.e., specific) binding was 3.2 nmol/mg of membrane protein. Oleate-agarose affinity chromatography of Triton X-100-solubilized LPM was used to isolate a single 40-kDa protein with high affinity for oleate. On gel filtration, the protein comigrated with various fatty acids but not with [14C]bilirubin, [35S]sulfobromophthalein, [14C]taurocholate, [14C]phosphatidylcholine, or [14C]cholesteryloleate. A rabbit antibody to this membrane fatty acid-binding protein gave a single precipitin line with the antigen but no reactivity with concentrated cytosolic proteins, LPM bilirubin/sulfobromophthalein-binding protein, or rat albumin or other rat plasma proteins. The antibody selectively inhibited heat-sensitive binding of [14C]oleate to LPM. Immunofluorescence studies localized the antigen in liver-cell plasma membranes as well as in other major sites of fatty acid transport. These data are compatible with the hypothesis that this protein may act as a receptor in a hepatocellular uptake mechanism for fatty acids.

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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. Accatino L., Simon F. R. Identification and characterization of a bile acid receptor in isolated liver surface membranes. J Clin Invest. 1976 Feb;57(2):496–508. doi: 10.1172/JCI108302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker K. J., Bradley S. E. Binding of sulfobromophthalein (BSP) sodium by plasma albumin. Its role in hepatic BSP extraction. J Clin Invest. 1966 Feb;45(2):281–287. doi: 10.1172/JCI105341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barrett P. V., Mullins F. X., Berlin N. I. Studies on the biosynthesis production of bilirubin-C14: an improved method utilizing delta-aminolevulinic acid-4-C14 in dogs. J Lab Clin Med. 1966 Dec;68(6):905–912. [PubMed] [Google Scholar]
  5. 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]
  6. Chen R. F. Removal of fatty acids from serum albumin by charcoal treatment. J Biol Chem. 1967 Jan 25;242(2):173–181. [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Dewald B., Dulaney J. T., Touster O. Solubilization and polyacrylamide gel electrophoresis of membrane enzymes with detergents. Methods Enzymol. 1974;32:82–91. doi: 10.1016/0076-6879(74)32011-3. [DOI] [PubMed] [Google Scholar]
  10. Erlinger S. Hepatocellular uptake of taurocholate in the dog. J Clin Invest. 1975 Feb;55(2):419–426. doi: 10.1172/JCI107946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fisher M. M., Bloxam D. L., Oda M., Phillips M. J., Yousef I. M. Characterization of rat liver cell plasma membranes. Proc Soc Exp Biol Med. 1975 Oct;150(1):177–184. doi: 10.3181/00379727-150-38998. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Kramer W., Buscher H. P., Gerok W., Kurz G. Bile salt binding to serum components. Taurocholate incorporation into high-density lipoprotein revealed by photoaffinity labelling. Eur J Biochem. 1979 Dec;102(1):1–9. doi: 10.1111/j.1432-1033.1979.tb06257.x. [DOI] [PubMed] [Google Scholar]
  14. Kushlan M. C., Gollan J. L., Ma W. L., Ockner R. K. Sex differences in hepatic uptake of long chain fatty acids in single-pass perfused rat liver. J Lipid Res. 1981 Mar;22(3):431–436. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Levi A. J., Gatmaitan Z., Arias I. M. Two hepatic cytoplasmic protein fractions, Y and Z, and their possible role in the hepatic uptake of bilirubin, sulfobromophthalein, and other anions. J Clin Invest. 1969 Nov;48(11):2156–2167. doi: 10.1172/JCI106182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lunazzi G., Tiribelli C., Gazzin B., Sottocasa G. Further studies on bilitranslocase, a plasma membrane protein involved in hepatic organic anion uptake. Biochim Biophys Acta. 1982 Feb 23;685(2):117–122. doi: 10.1016/0005-2736(82)90087-6. [DOI] [PubMed] [Google Scholar]
  18. Mahadevan S., Sauer F. Effect of trypsin, phospholipases, and membrane-impermeable reagents on the uptake of palmitic acid by isolated rat liver cells. Arch Biochem Biophys. 1974 Sep;164(1):185–193. doi: 10.1016/0003-9861(74)90021-6. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. OSTROW J. D., SCHMID R. THE PROTEIN-BINDING OF C14-BILIRUBIN IN HUMAN AND MURINE SERUM. J Clin Invest. 1963 Aug;42:1286–1299. doi: 10.1172/JCI104813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. OUCHTERLONY O. Diffusion-in-gel methods for immunological analysis. Prog Allergy. 1958;5:1–78. [PubMed] [Google Scholar]
  22. Ockner R. K., Manning J. A. Fatty acid-binding protein in small intestine. Identification, isolation, and evidence for its role in cellular fatty acid transport. J Clin Invest. 1974 Aug;54(2):326–338. doi: 10.1172/JCI107768. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Paris S., Samuel D., Romey G., Ailhaud G. Uptake of fatty acids by cultured cardiac cells from chick embryo: evidence for a facilitation process without energy dependence. Biochimie. 1979;61(3):361–367. doi: 10.1016/s0300-9084(79)80129-7. [DOI] [PubMed] [Google Scholar]
  24. Paumgartner G., Reichen J. Different pathways for hepatic uptake of taurocholate and indocyanine green. Experientia. 1975 Mar 15;31(3):306–308. doi: 10.1007/BF01922553. [DOI] [PubMed] [Google Scholar]
  25. Paumgartner G., Reichen J. Kinetics of hepatic uptake of unconjugated bilirubin. Clin Sci Mol Med. 1976 Aug;51(2):169–176. doi: 10.1042/cs0510169. [DOI] [PubMed] [Google Scholar]
  26. Peters T., Jr, Taniuchi H., Anfinsen C. B., Jr Affinity chromatography of serum albumin with fatty acids immobilized on agarose. J Biol Chem. 1973 Apr 10;248(7):2447–2451. [PubMed] [Google Scholar]
  27. Porath J., Sundberg L. High capacity chemisorbents for protein immobilization. Nat New Biol. 1972 Aug 30;238(87):261–262. doi: 10.1038/newbio238261a0. [DOI] [PubMed] [Google Scholar]
  28. Reichen J., Berk P. D. Isolation of an organic anion binding protein from rat liver plasma membrane fractions by affinity chromatography. Biochem Biophys Res Commun. 1979 Nov 28;91(2):484–489. doi: 10.1016/0006-291x(79)91547-x. [DOI] [PubMed] [Google Scholar]
  29. Reichen J., Blitzer B. L., Berk P. D. Binding of unconjugated and conjugated sulfobromophthalein to rat liver plasma membrane fractions in vitro. Biochim Biophys Acta. 1981 Jan 8;640(1):298–312. doi: 10.1016/0005-2736(81)90554-x. [DOI] [PubMed] [Google Scholar]
  30. Reichen J., Paumgartner G. Kinetics of taurocholate uptake by the perfused rat liver. Gastroenterology. 1975 Jan;68(1):132–136. [PubMed] [Google Scholar]
  31. 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]
  32. Scharschmidt B. F., Waggoner J. G., Berk P. D. Hepatic organic anion uptake in the rat. J Clin Invest. 1975 Nov;56(5):1280–1292. doi: 10.1172/JCI108204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Stremmel W., Gerber M. A., Glezerov V., Thung S. N., Kochwa S., Berk P. D. Physicochemical and immunohistological studies of a sulfobromophthalein- and bilirubin-binding protein from rat liver plasma membranes. J Clin Invest. 1983 Jun;71(6):1796–1805. doi: 10.1172/JCI110935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Stremmel W., Kochwa S., Berk P. D. Studies of oleate binding to rat liver plasma membranes. Biochem Biophys Res Commun. 1983 Apr 15;112(1):88–95. doi: 10.1016/0006-291x(83)91801-6. [DOI] [PubMed] [Google Scholar]
  35. Stremmel W., Potter B. J., Berk P. D. Studies of albumin binding to rat liver plasma membranes. Implications for the albumin receptor hypothesis. Biochim Biophys Acta. 1983 Mar 15;756(1):20–27. doi: 10.1016/0304-4165(83)90019-3. [DOI] [PubMed] [Google Scholar]
  36. Wolkoff A. W., Chung C. T. Identification, purification, and partial characterization of an organic anion binding protein from rat liver cell plasma membrane. J Clin Invest. 1980 May;65(5):1152–1161. doi: 10.1172/JCI109770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. von Dippe P., Levy D. Characterization of the bile acid transport system in normal and transformed hepatocytes. Photoaffinity labeling of the taurocholate carrier protein. J Biol Chem. 1983 Jul 25;258(14):8896–8901. [PubMed] [Google Scholar]

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