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Characterization and binding properties of human fetal lung fatty acid-binding proteins

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Abstract

When delipidated Mr>10,000 cut-off human fetal lung cytosol was separated on gel filtration and ion-exchange chromatography on Auto-FPLC system, two fatty acid-binding proteins (FABPs) of pI 6.9 and pI 5.4 were purified to homogeneity. On Western blotting analysis with the anti-human fetal lung pI 6.9 FABP, these two proteins showed immunochemical cross reactivity with each other and with purified hepatic FABPs but not with cardiac or gut FABP. These two FABPs have identical molecular mass of 15.2 kDa, which is slightly higher than that of the hepatic proteins (14.2 kDa). Carbohydrate covalently linked to FABPs, that may substantially add to the molecular mass, was not detected in the purified protein preparations. Amino acid analysis revealed that both the proteins have same amino acid composition each containing one Trp residue that is lacking in hepatic FABP. Different isoforms of lung FABP exhibited different binding ability for their natural ligands. These proteins bind palmitoyl CoA with higher affinity than oleic acid. pI 6.9 FABP can more rapidly and efficiently transfer fatty acid than can pI 5.4 FABP from unilammelar liposomes. Thus these FABPs may play a critical role in fatty acid transport during human fetal lung development.

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Abbreviations

AO:

anthroyloxy

12-AS:

12-(9-anthroyloxy)stearic acid

FABP:

fatty acid-binding protein

NBD-PE:

[N-(4-nitrobenzo-2-oxa-1,3-diazole)phosphatidylethanolamine

Pal-CoA:

palmitoyl coenzyme A

PITC:

phenylisothiocyanate

PBS:

phosphate-buffered saline

PtdCho:

phosphatidylcholine

SUV:

small unilamellar vesicle

Tris:

tris(hydroxymethyl) amino methane

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Authors and Affiliations

  1. Department of Biochemistry, University of Calcutta, College of Science, 35 Ballygunge Circular Road, 700 019, Calcutta, India

    Tanya Das & Manju Mukherjea

  2. Department of Cell Biology, Cleveland Clinic Foundation Research Institute, 9500 Euclid Avenue, 44195, Cleveland, Ohio, USA

    Gaurisankar Sa

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Sa, G., Das, T. & Mukherjea, M. Characterization and binding properties of human fetal lung fatty acid-binding proteins. Mol Cell Biochem 129, 67–75 (1993). https://doi.org/10.1007/BF00926577

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  • DOI: https://doi.org/10.1007/BF00926577

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