title: Expression, purification and functional characterization of recombinant human acyl-CoA-binding protein (ACBP) from erythroid cells.
abstract: 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.
authors.affiliation: Department of Gastrointestinal and General Surgery, Wroclaw Medical University, Poland. augoffka@chir.am.wroc.pl
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Augoff
authors.forename: Adam
authors.initials: A
authors.lastname: Kolondra
authors.forename: Anna
authors.initials: A
authors.lastname: Chorzalska
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Lach
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Grabowski
authors.forename: Aleksander F
authors.initials: AF
authors.lastname: Sikorski
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D026261:Diazepam Binding Inhibitor / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D041905:Erythroid Cells / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D006801:Humans; D054878:Lipoylation / Q000187:drug effects; D008969:Molecular Sequence Data; D011485:Protein Binding; D020033:Protein Isoforms; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D026261:Diazepam Binding Inhibitor; D020033:Protein Isoforms; D011994:Recombinant Proteins