Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Rolf, B; Oudenampsen-Krüger, E; Börchers, T; Faergeman, NJ; Knudsen, J; Lezius, A; Spener, F.
Analysis of the ligand binding properties of recombinant bovine liver-type fatty acid binding protein.
Biochim Biophys Acta. 1995; 1259(3):245-253
Doi: 10.1016/0005-2760(95)00170-0
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- Co-Autor*innen der Med Uni Graz
- Spener Friedrich
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- Abstract:
- The coding part of the cDNA for bovine liver-type fatty acid binding protein (L-FABP) has been amplified by RT-PCR, cloned and used for the construction of an Escherichia coli (E. coli) expression system. The recombinant protein made up to 25% of the soluble E. coli proteins and could be isolated by a simple two step protocol combining ion exchange chromatography and gel filtration. Dissociation constants for binding of oleic acid, arachidonic acid, oleoyl-CoA, lysophosphatidic acid and the peroxisomal proliferator bezafibrate to L-FABP have been determined by titration calorimetry. All ligands were bound in a 2:1 stoichiometry, the dissociation constants for the first ligand bound were all in the micro molar range. Oleic acid was bound with the highest affinity and a Kd of 0.26 microM. Furthermore, binding of cholesterol to L-FABP was investigated with the Lipidex assay, a liposome binding assay and a fluorescence displacement assay. In none of the assays binding of cholesterol to L-FABP was observed.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Animals -
- Base Sequence -
- Calorimetry -
- Carrier Proteins - chemistry Carrier Proteins - genetics Carrier Proteins - metabolism
- Cattle -
- Cholesterol - metabolism
- Cloning, Molecular -
- Dextrans -
- Fatty Acid-Binding Proteins -
- Fatty Acids - chemistry Fatty Acids - metabolism
- Fluorescence -
- Ligands -
- Liposomes - chemistry
- Liver - metabolism
- Lysophospholipids - metabolism
- Molecular Sequence Data -
- Myelin P2 Protein - chemistry Myelin P2 Protein - genetics Myelin P2 Protein - metabolism
- Neoplasm Proteins -
- Recombinant Proteins - chemistry Recombinant Proteins - metabolism
- Sequence Analysis -
- Thermodynamics -
- Find related publications in this database (Keywords)
- liver-type fatty acid binding protein
- titration calorimetry
- ligand binding