High-yield two-step chromatographic procedure for purification of fatty acid-binding protein from human heart
| Autor: | Ger J. van der Vusse, M. M. Vork, Frans A. van Nieuwenhoven, Don A. M. Surtel, Jan F. C. Glatz, Appie H. Kleine |
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| Rok vydání: | 1991 |
| Předmět: |
Fatty Acid-Binding Proteins
Sepharose Gel permeation chromatography chemistry.chemical_compound Humans Polyacrylamide gel electrophoresis chemistry.chemical_classification Gel electrophoresis Chromatography Isoelectric focusing Myocardium Tumor Suppressor Proteins Fatty Acids Reproducibility of Results Fatty acid General Chemistry Chromatography Ion Exchange Neoplasm Proteins Dissociation constant Oleic acid chemistry Chromatography Gel Electrophoresis Polyacrylamide Gel Spectrophotometry Ultraviolet lipids (amino acids peptides and proteins) Isoelectric Focusing Carrier Proteins Fatty Acid-Binding Protein 7 |
| Zdroj: | Journal of Chromatography B: Biomedical Sciences and Applications. 570:173-179 |
| ISSN: | 0378-4347 |
| Popis: | A high-yield procedure for the purification of cytoplasmic fatty acid-binding protein from human heart (H-FABP) is described. H-FABP was purified by gel permeation chromatography on a Sephacryl S-200 column followed by anion-exchange chromatography on a Sepharose Q fast-flow column at pH 7.0. At this pH H-FABP binds strongly to the column and can be selectively eluted with a salt gradient. The two-step procedure showed a high degree of reproducibility. On average 50 mg of H-FABP was obtained from 150 g of human heart tissue, which corresponds to a recovery of about 50%. Purity was confirmed by gel electrophoresis and isoelectric focusing. Binding of oleic acid to purified H-FABP, using the Lipidex 1000 assay, revealed a maximal binding of 0.75 +/- 0.01 mol fatty acid/mol protein and a dissociation constant of 0.19 +/- 0.01 microM. |
| Databáze: | OpenAIRE |
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