Studies on fatty acid-binding proteins. The diurnal variation shown by rat liver fatty acid-binding protein

The concentration of fatty acid-binding protein in rat liver was examined by SDS/polyacrylamide-gel electrophoresis, by Western blotting and by quantifying the fluorescence enhancement achieved on the binding of the fluorescent probe 11-(dansylamino)undecanoic acid. A 2-3-fold increase in the concentration of this protein produced by treatment of rats with the peroxisome proliferator tiadenol was readily detected; however, only a small variation in the concentration of the protein due to a diurnal rhythm was observed. This result contradicts the 7-10-fold variation previously reported for this protein [Hargis, Olson, Clarke & Dempsey (1986) J. Biol. Chem. 261, 1988-1991].

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

Biochemical Journal ◽

10.1042/bj2380419 ◽

1986 ◽

Vol 238 (2) ◽

pp. 419-424 ◽

Cited By ~ 48

Author(s):

T C I Wilkinson ◽

D C Wilton

Keyword(s):

Fatty Acid ◽

Rat Liver ◽

Fatty Acid Binding Protein ◽

Binding Protein ◽

Fluorescence Emission ◽

Fatty Acid Binding Proteins ◽

Liver Fatty Acid ◽

Fatty Acid Binding ◽

Acid Binding Protein ◽

Acid Analogue

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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Studies on fatty-acid-binding proteins. The purification of rat liver fatty-acid-binding protein and the role of cysteine-69 in fatty acid binding

Biochemical Journal ◽

10.1042/bj2610273 ◽

1989 ◽

Vol 261 (1) ◽

pp. 273-276 ◽

Cited By ~ 23

Author(s):

D C Wilton

Keyword(s):

Fatty Acid ◽

Rat Liver ◽

Fatty Acid Binding Protein ◽

Binding Protein ◽

Cysteine Residue ◽

High Yield ◽

Fatty Acid Binding Proteins ◽

Affinity Column ◽

Fatty Acid Binding ◽

Acid Binding Protein

1. A new, simple and high-yield procedure is described for the purification of hepatic fatty-acid-binding protein from rat liver using naphthylaminodecyl-agarose as an affinity column. 2. Cysteine-69 is shown to react slowly, but quantitatively, with 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB), indicating that the thiol group is free, but may be buried within the protein. 3. Fatty acids do not affect the DTNB reactivity of this cysteine residue; however, cysteine reactivity is enhanced in the presence of haem and oleoyl-CoA. 4. Fatty-acid-binding protein that has been modified with DTNB is still able to bind the fluorescent fatty acid 11-(dansylamino)undecanoic acid, indicating that cysteine-69 may be remote from the fatty-acid-binding site.

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