期刊
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR AND CELL BIOLOGY OF LIPIDS
卷 1488, 期 3, 页码 245-254出版社
ELSEVIER
DOI: 10.1016/S1388-1981(00)00128-1
关键词
adipose differentiation related protein; fatty acid binding protein; fatty acid; adipocyte; lipid
资金
- NIDDK NIH HHS [DK 51463, DK 401402] Funding Source: Medline
Adipose differentiation related protein (ADRP) is a 53 kDa protein encoded by a cDNA originally cloned by differential hybridization from murine adipocytes. ADRP is induced during the early onset of the adipose differentiation program and is expressed at high level in mature adipocytes. We have demonstrated that ADRP stimulated the uptake of fatty acids thereby providing evidence for a functional role of ADRP in lipid metabolism. Tn the present paper, the murine ADRP has been expressed as a recombinant histidine-tagged protein in Escherichia coli, and purified from expressing cultures in order to examine its biochemical properties. We report here that the purified recombinant ADRP binds fatty acids and exhibits stoichiometric saturable binding of NBD-stearic acid with a K-d = 0.145 +/- 0.003 muM and a B-max = 0.99 +/- 0.05. Analysis of fluorescence emission spectra indicates that the polarity of the ADRP binding site is near epsilon approximate to 23, close to that observed for fatty acid binding sites in other lipid binding proteins such as the liver fatty acid binding protein. The data presented here provide evidence that isolated ADRP purified in the experimental conditions described hen call be used for functional studies. (C) 2000 Elsevier Science B.V. All rights reserved.
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