Expression, purification and functional characterization of recombinant human acyl-CoA-binding protein (ACBP) from erythroid cells
| Autor: | Krzysztof Grabowski, Anna Chorzalska, Aleksander F. Sikorski, Katarzyna Augoff, Agnieszka Lach, Adam Kolondra |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Lipoylation
Molecular Sequence Data Cellular homeostasis Biology General Biochemistry Genetics and Molecular Biology law.invention chemistry.chemical_compound Protein acylation Erythroid Cells Palmitoylation law Phosphatidylcholine Acyl-CoA-binding protein Escherichia coli Humans Protein Isoforms Protein palmitoylation Amino Acid Sequence Cloning Molecular Diazepam Binding Inhibitor Recombinant Proteins Gene Expression Regulation Biochemistry chemistry Recombinant DNA lipids (amino acids peptides and proteins) Intracellular Protein Binding |
| Zdroj: | National Information Processing Institute Scopus-Elsevier |
| ISSN: | 1734-154X 0001-527X |
| DOI: | 10.18388/abp.2010_2440 |
| Popis: | Fatty acyl-CoA esters are extremely important in cellular homeostasis. They are intermediates in both lipid metabolism and post-translational protein modifications. Among these modification events, protein palmitoylation seems to be unique by its reversibility which allows dynamic regulation of the protein hydrophobicity. The recent discovery of an enzyme family that catalyze protein palmitoylation has increased the understanding of the enzymology of the covalent attachment of fatty acids to proteins. Despite that, the molecular mechanism of supplying acyl-CoA esters to this reaction is yet to be established. Acyl-coenzyme A-binding proteins are known to bind long-chain acyl-CoA esters with very high affinity. Therefore, they play a significant role in intracellular acyl-CoA transport and pool formation. The purpose of this work is to explore the potential of one of the acyl-CoA-binding proteins to participate in the protein palmitoylation. In this study, a recombinant form of ACBP derived from human erythroid cells was expressed in E. coli, purified, and functionally characterized. We demonstrate that recombinant hACBP effectively binds palmitoyl-CoA in vitro, undergoing a shift from a monomeric to a dimeric state, and that this ligand-binding ability is involved in erythrocytic membrane phosphatidylcholine (PC) remodeling but not in protein acylation. |
| Databáze: | OpenAIRE |
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