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. 1986 Sep 1;238(2):419–424. doi: 10.1042/bj2380419

Studies on fatty acid-binding proteins. The detection and quantification of the protein from rat liver by using a fluorescent fatty acid analogue.

T C Wilkinson, D C Wilton
PMCID: PMC1147152  PMID: 3800946

Abstract

Fatty acid-binding protein from rat liver is shown to bind the fluorescent fatty acid probe dansyl undecanoic acid. Binding is accompanied by a shift in the fluorescence emission maximum from 550 nm to 500 nm and a 60-fold fluorescence enhancement at 500 nm. These spectral properties have allowed the use of this probe to detect and quantify microgram amounts of liver fatty acid-binding protein during purification procedures. In conjunction with h.p.l.c. the method allows the rapid estimation of liver fatty acid-binding protein in biological samples. The validity of the method is demonstrated by measuring the concentration of fatty acid-binding protein in livers from control and hypolipidaemic-drug-treated rats. The dramatic diurnal rhythm previously reported for this protein [Dempsey (1984) Curr. Top. Cell. Regul. 24, 63-86] was not observed with this method.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bass N. M., Manning J. A., Ockner R. K., Gordon J. I., Seetharam S., Alpers D. H. Regulation of the biosynthesis of two distinct fatty acid-binding proteins in rat liver and intestine. Influences of sex difference and of clofibrate. J Biol Chem. 1985 Feb 10;260(3):1432–1436. [PubMed] [Google Scholar]
  2. Bass N. M., Manning J. A., Ockner R. K. Turnover and short-term regulation of fatty acid binding protein in liver. J Biol Chem. 1985 Aug 15;260(17):9603–9607. [PubMed] [Google Scholar]
  3. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  4. Dempsey M. E., McCoy K. E., Baker H. N., Dimitriadou-Vafiadou A., Lorsbach T., Howard J. B. Large scale purification and structural characterization of squalene and sterol carrier protein. J Biol Chem. 1981 Feb 25;256(4):1867–1873. [PubMed] [Google Scholar]
  5. Dempsey M. E. Regulation of lipid metabolism by a lipid-carrying protein. Curr Top Cell Regul. 1984;24:63–86. doi: 10.1016/b978-0-12-152824-9.50014-9. [DOI] [PubMed] [Google Scholar]
  6. Dottavio-Martin D., Ravel J. M. Radiolabeling of proteins by reductive alkylation with [14C]formaldehyde and sodium cyanoborohydride. Anal Biochem. 1978 Jul 1;87(2):562–565. doi: 10.1016/0003-2697(78)90706-6. [DOI] [PubMed] [Google Scholar]
  7. Fleischner G., Meijer D. K., Levine W. G., Gatmaitan Z., Gluck R., Arias I. M. Effect of hypolipidemic drugs, nafenopin and clofibrate, on the concentration of ligandin and Z protein in rat liver. Biochem Biophys Res Commun. 1975 Dec 15;67(4):1401–1407. doi: 10.1016/0006-291x(75)90182-5. [DOI] [PubMed] [Google Scholar]
  8. Glatz J. F., Baerwaldt C. C., Veerkamp J. H., Kempen H. J. Diurnal variation of cytosolic fatty acid-binding protein content and of palmitate oxidation in rat liver and heart. J Biol Chem. 1984 Apr 10;259(7):4295–4300. [PubMed] [Google Scholar]
  9. 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]
  10. Glatz J. F., Veerkamp J. H. A radiochemical procedure for the assay of fatty acid binding by proteins. Anal Biochem. 1983 Jul 1;132(1):89–95. doi: 10.1016/0003-2697(83)90429-3. [DOI] [PubMed] [Google Scholar]
  11. Glatz J. F., Veerkamp J. H. Intracellular fatty acid-binding proteins. Int J Biochem. 1985;17(1):13–22. doi: 10.1016/0020-711x(85)90080-1. [DOI] [PubMed] [Google Scholar]
  12. Grinstead G. F., Trzaskos J. M., Billheimer J. T., Gaylor J. L. Cytosolic modulators of activities of microsomal enzymes of cholesterol biosynthesis. Effects of Acyl-CoA inhibition and cytosolic Z-protein. Biochim Biophys Acta. 1983 Mar 22;751(1):41–51. doi: 10.1016/0005-2760(83)90255-2. [DOI] [PubMed] [Google Scholar]
  13. Horowitz P. M., Criscimagna N. L. Differential binding of the fluorescent probe 8-anilinonaphthalene-2-sulfonic acid to rhodanese catalytic intermediates. Biochemistry. 1985 May 21;24(11):2587–2593. doi: 10.1021/bi00332a001. [DOI] [PubMed] [Google Scholar]
  14. Kawashima Y., Nakagawa S., Tachibana Y., Kozuka H. Effects of peroxisome proliferators on fatty acid-binding protein in rat liver. Biochim Biophys Acta. 1983 Nov 1;754(1):21–27. doi: 10.1016/0005-2760(83)90077-2. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Morrow F. D., Martin R. J. Quantitation of hepatic fatty acid-binding proteins by post-chromatographic ligand binding assay. J Lipid Res. 1983 Mar;24(3):324–331. [PubMed] [Google Scholar]
  17. Ockner R. K., Burnett D. A., Lysenko N., Manning J. A. Sex differences in long chain fatty acid utilization and fatty acid binding protein concentration in rat liver. J Clin Invest. 1979 Jul;64(1):172–181. doi: 10.1172/JCI109437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ockner R. K., Manning J. A., Kane J. P. Fatty acid binding protein. Isolation from rat liver, characterization, and immunochemical quantification. J Biol Chem. 1982 Jul 10;257(13):7872–7878. [PubMed] [Google Scholar]
  19. Renaud G., Foliot A., Infante R. Increased uptake of fatty acids by the isolated rat liver after raising the fatty acid binding protein concentration with clofibrate. Biochem Biophys Res Commun. 1978 Jan 30;80(2):327–334. doi: 10.1016/0006-291x(78)90680-0. [DOI] [PubMed] [Google Scholar]
  20. Takahashi K., Odani S., Ono T. Isolation and characterization of the three fractions (DE-I, DE-II and DE-III) of rat-liver Z-protein and the complete primary structure of DE-II. Eur J Biochem. 1983 Nov 15;136(3):589–601. doi: 10.1111/j.1432-1033.1983.tb07781.x. [DOI] [PubMed] [Google Scholar]
  21. Whitaker J. R., Granum P. E. An absolute method for protein determination based on difference in absorbance at 235 and 280 nm. Anal Biochem. 1980 Nov 15;109(1):156–159. doi: 10.1016/0003-2697(80)90024-x. [DOI] [PubMed] [Google Scholar]

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