Expression, purification and functional characterization of recombinant human acyl-CoA-binding protein (ACBP) from erythroid cells

• Authors: Katarzyna Augoff , Adam Kolondra , Anna Chorzalska , Agnieszka Lach , Krzysztof Grabowski , Aleksander F Sikorski
• Languages of publication: EN

Abstracts

EN

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.

Keywords

EN

acyl-CoA-binding protein; PC; DBI; ACBP; phosphatidylcholine

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2010
• Volume: 57
• Issue: 4
• Pages: 533-540

Dates

published

2010

received

2010-04-19

revised

2010-10-06

accepted

2010-11-13

(unknown)

2010-11-16

Contributors

author

Katarzyna Augoff

• Department of Gastrointestinal and General Surgery, Wroclaw Medical University, Wroclaw, Poland

author

Adam Kolondra

• Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wroclaw, Wroclaw, Poland

author

Anna Chorzalska

• Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wroclaw, Wroclaw, Poland

author

Agnieszka Lach

• Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wroclaw, Wroclaw, Poland

author

Krzysztof Grabowski

• Department of Gastrointestinal and General Surgery, Wroclaw Medical University, Wroclaw, Poland

author

Aleksander F Sikorski

• Laboratory of Cytobiochemistry, Biotechnology Faculty, University of Wroclaw, Wroclaw, Poland

References

• Abo-Hashema KAH, Cake MH, Lukas MA, Knudsen J (2001) The interaction of acyl-CoA with acyl-CoA binding protein and carnitine palmitoyltransferase I. Int J Biochem Cell Biol 33: 807-815.
• Baekkeskov N, Kanaani J (2009) Palmitoylation cycles and regulation of protein function. Mol Membr Biol 26: 42-54.
• Boylan JG, Hamilton JA (1992) Interactions of acyl-Coenzyme A with phosphatidylcholine bilayers and serum albumin? Biochemistry 31: 557-567.
• Dise CA, Goodman DBP, Rasmussen H (1980) Definition of the pathway for membrane phospholipid fatty acid turnover in human erythrocytes. J Lipid Res 21: 292-300.
• Dodge JT, Mitchell C, Hanahan DJ (1963) The preparation and chemical characteristics of hemoglobin-free ghosts of erythrocytes. Arch Biochem 100: 119-130.
• Dunphy JT, Schroeder H, Leventis R, Greentree WK, Knudsen JK, Silvius JR, Linder ME (2000) Differential effects of acyl-CoA binding protein on enzymatic and non-enzymatic thioacylation of protein and peptide substrates. Biochim Biophys Acta 1485: 185-198.
• Færgeman NJ, Knudsen J (1997) Role of long-chain fatty acyl-CoA esters in the regulation of metabolism and in cell signaling. Biochem J 323: 1-12.
• Færgeman NJ, Wadum M, Feddersen S, Burton M, Kragelund BB, Knudsen J (2007) Acyl-CoA binding proteins; structural and functional conservation over 2000 MYA. Mol Cell Biochem 299: 55-65.
• Fyrst H, Knudsen J, Schott MA, Lubin BH, Kuypers FA (1995) Detection of acyl-CoA-binding protein in human red blood cells and investigation of its role in membrane phospholipid renewal. Biochem J 306: 793-799.
• Gossett RE, Frolov AA, Roths JB, Behnke WD, Kier AB, Schroeder F (1996) Acyl-CoA binding proteins: multiplicity and function. Lipids 31: 895-918.
• Gossett RE, Edmondson RD, Jolly CA, Cho TH, Russell DH, Knudsen J, Kier AB, Schroeder F (1998) Structure and function of normal and transformed murine Acyl-CoA binding proteins. Arch Biochem Biophys 350: 201-213.
• Guidotti A, Forchetti CM, Corda MG, Konkel D, Bennett DD, Costa E (1983) Isolation, characterization, and purification to homogeneity of an endogenous polypeptide with agonistic action on benzodiazepine receptors (endogenous benzodiazepine ligand/GABA receptor/anxiety/partial peptide sequence). Proc Natl Acad Sci U S A 80: 3531-3535.
• Hu L, Deeney JT, Nolan CJ, Peyot ML, Ao A, Richard AM, Luc E, Faergeman NJ, Knudsen J, Guo W, Sorhede-Winzell M, Prentki M, Corkey BE (2005) Regulation of lipolytic activity by long-chain acyl-coenzyme A in islets and adipocytes. Am J Physiol Endocrinol Metab 289: E1085-E1092.
• Iwanaga T, Tsutsumi R, Noritake J, Fukata Y, Fukata M (2009) Dynamic protein palmitoylation in cellular signaling. Prog Lipid Res 48: 117-127.
• Jolly CA, Wilton DC, Schroeder F (2000) Microsomal fatty acyl-CoA transacylation and hydrolysis: fatty acyl-CoA species dependent modulation by liver fatty acyl-CoA binding proteins. Biochim Biophys Acta 1483: 185-197.
• Knudsen J, Højrup P, Hansen HO, Hansen HF, Roepstroff P (1989) Acyl-CoA-binding protein in the rat. Purification, binding characteristics, tissue concentrations and amino acid sequence. Biochem J 262: 513-519.
• Knudsen J, Færgeman NJ, Skøtt H, Hummel R, Børsting C, Rose TM, Andersen JS, Højrup P, Roepstorfft P, Kristiansen K (1994) Yeast acyl-CoA-binding protein: acyl-CoA-binding affinity and effect on intracellular acyl-CoA pool size. Biochem J 302: 479-485.
• Knudsen J, Neergaard TBF, Gaigg B, Jensen MV, Hansen JK (2000) Role of acyl-CoA binding protein in acyl-CoA metabolism and acyl-CoA-mediated cell signaling. J Nutr 130: 294S-298S.
• Leventis R, Juel G, Knudsen JK, Silvius JR (1987) Acyl-CoA binding proteins inhibit the nonenzymic S-acylation of cysteinyl-containing peptide sequences by long-chain acyl-CoAs. Biochemistry 36: 5546-5553.
• Linder ME, Deschenes RJ (2007) Palmitoylation: policing protein stability and traffic. Nature 8: 74-84.
• Mitchell GA, Gauthier N, Lesimple A, Wang SP, Mamer O, Qureshi I (2008) Hereditary and acquired diseases of acyl-coenzyme A metabolism. Mol Genet Metab 94: 4-15.
• Monzani PS, Pereira HM, Melo FA, Meirelles FV, Oliva G, Cascardo JCM (2010) A new topology of ACBP from Moniliophthora perniciosa. Biochim Biophys Acta 1804: 115-123.
• Pfanner N, Glick BS, Arden SR, Rothman JE (1990) Fatty acylation promotes fusion of transport vesicles with golgi cisternae. J Cell Biol 110: 955-961.
• Rasmussen JT, Fiergeman NJ, Kristiansent K, Knudsen J (1994) Acyl-CoA-binding protein (ACBP) can mediate intermembrane acyl-CoA transport and donate acyl-CoA for b-oxidation and glycerolipid synthesis. Biochem J 299: 165-170.
• Rosendal J, Ertbjerg P, Knudsen J (1993) Characterization of ligand binding to acyl-CoA-binding protein. Biochem J 290: 321-326.
• Schmidt MFG, McIlhinney RAJ, Burns GR (1995) Palmitoylation of endogenous and viral acceptor proteins by fatty acyltransferase (PAT) present in erythrocyte ghosts and in placental membranes. Biochim Biophys Acta 1257: 205-213.
• Shindou H, Shimi T (2009) Acyl-CoA:lysophospholipid acyltransferases. J Biol Chem 284: 1-5.
• Shrago E (2000) Long-chain acyl-CoA as a multi-effector ligand in cellular metabolism. J Nutr 130: 290S-293S.
• Simonsen AC, Jensen UB, Færgeman NJ, Knudsen J, Mouritsen OG (2003) Acyl-coenzyme A organizes laterally in membranes and is recognized specifically by acyl-coenzyme A binding protein. FEBS Lett 552: 253-258.
• Soupene E, Fyrst H, Kuypers FA (2008) Mammalian acyl-CoA:lysophosphatidylcholine acyltransferase enzymes. Proc Natl Acad Sci USA 105: 88-93.
• Taskinen JP, van Aalten DM, Knudsen J, Wierenga RK (2007) High resolution crystal structures of unliganded and liganded human liver ACBP reveal a new mode of binding for the acyl-CoA ligand. Proteins: Structure, Function and Bioinformatics 66: 229-238.