In the present study, we microinjected fluorescently labelled liver bovine ACBP (acyl-CoA-binding protein) [FACI-50 (fluorescent acyl-CoA indicator-50)] into HeLa and BMGE (bovine mammary gland epithelial) cell lines to characterize the localization and dynamics of ACBP in living cells. Results showed that ACBP targeted to the ER (endoplasmic reticulum) and Golgi in a ligand-binding-dependent manner. A variant Y28F/K32A-FACI-50, which is unable to bind acyl-CoA, did no longer show association with the ER and became segregated from the Golgi, as analysed by intensity correlation calculations. Depletion of fatty acids from cells by addition of FAFBSA (fatty-acid-free BSA) significantly decreased FACI-50 association with the Golgi, whereas fatty acid overloading increased Golgi association, strongly supporting that ACBP associates with the Golgi in a ligand-dependent manner. FRAP (fluorescence recovery after photobleaching) showed that the fatty-acid-induced targeting of FACI-50 to the Golgi resulted in a 5-fold reduction in FACI-50 mobility. We suggest that ACBP is targeted to the ER and Golgi in a ligand-binding-dependent manner in living cells and propose that ACBP may be involved in vesicular trafficking.
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Research Article|
February 27 2008
Acyl-CoA-binding protein (ACBP) localizes to the endoplasmic reticulum and Golgi in a ligand-dependent manner in mammalian cells
Jesper S. Hansen;
Jesper S. Hansen
*Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark
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Nils J. Færgeman;
Nils J. Færgeman
*Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark
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Birthe B. Kragelund;
Birthe B. Kragelund
†Department of Molecular Biology, Ole Maaløes Vej 5, DK-2200 Copenhagen N, Denmark
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Jens Knudsen
Jens Knudsen
1
*Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, DK-5230 Odense M, Denmark
1To whom correspondence should be addressed (email jkk@bmb.sdu.dk).
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Publisher: Portland Press Ltd
Received:
April 25 2007
Revision Received:
October 04 2007
Accepted:
October 22 2007
Accepted Manuscript online:
October 22 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biochem J (2008) 410 (3): 463–472.
Article history
Received:
April 25 2007
Revision Received:
October 04 2007
Accepted:
October 22 2007
Accepted Manuscript online:
October 22 2007
Citation
Jesper S. Hansen, Nils J. Færgeman, Birthe B. Kragelund, Jens Knudsen; Acyl-CoA-binding protein (ACBP) localizes to the endoplasmic reticulum and Golgi in a ligand-dependent manner in mammalian cells. Biochem J 15 March 2008; 410 (3): 463–472. doi: https://doi.org/10.1042/BJ20070559
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