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Brief Report

Intracellular Localization of Endogenous Mouse ABCG1 Is Mimicked by Both ABCG1-L550 and ABCG1-P550

Elizabeth J. Tarling, Peter A. Edwards
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https://doi.org/10.1161/ATVBAHA.116.307414
Arteriosclerosis, Thrombosis, and Vascular Biology. 2016;ATVBAHA.116.307414
Originally published May 26, 2016
Elizabeth J. Tarling
From the Departments of Biological Chemistry (P.A.E.) and Medicine (E.J.T.), David Geffen School of Medicine at the University of California, Los Angeles.
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Peter A. Edwards
From the Departments of Biological Chemistry (P.A.E.) and Medicine (E.J.T.), David Geffen School of Medicine at the University of California, Los Angeles.
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Abstract

Objective—In a recent article in Arteriosclerosis, Thrombosis, and Vascular Biology, it was reported that ATP-binding cassette transporter G1 (ABCG1) containing leucine at position 550 (ABCG1-L550) was localized to the plasma membrane, whereas ABCG1-P550 (proline at position 550) was intracellular. Because the published data on the subcellular localization of ABCG1 are controversial, we performed additional experiments to determine the importance of leucine or proline at amino acid 550.

Approach and Results—We transfected multiple cell lines (CHO-K1, Cos-7, and HEK293) with untagged or FLAG-tagged ABCG1 containing either leucine or proline at position 550. Immunofluorescence studies demonstrated that in all cases, ABCG1 localized to intracellular endosomal vesicles. We also show that both ABCG1-L550 and ABCG1-P550 are equally active in both promoting the efflux of cellular cholesterol to exogenous high-density lipoprotein and in inducing the activity of sterol regulatory element–binding protein-2, presumably as a result of redistributing intracellular sterols away from the endoplasmic reticulum. Importantly, we treated nontransfected primary peritoneal macrophages with a liver X receptor agonist and demonstrate, using immunofluorescence, that although endogenous ABCG1 localizes to intracellular endosomes, none was detectable at the cell surface/plasma membrane.

Conclusions—ABCG1, irrespective of either a leucine or proline at position 550, is an intracellular protein that localizes to vesicles of the endosomal pathway where it functions to mobilize sterols away from the endoplasmic reticulum and out of the cell.

  • ABC transporter
  • cholesterol homeostasis
  • lipids
  • Received February 17, 2016.
  • Accepted May 10, 2016.
  • © 2016 American Heart Association, Inc.
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    Intracellular Localization of Endogenous Mouse ABCG1 Is Mimicked by Both ABCG1-L550 and ABCG1-P550
    Elizabeth J. Tarling and Peter A. Edwards
    Arteriosclerosis, Thrombosis, and Vascular Biology. 2016;ATVBAHA.116.307414, originally published May 26, 2016
    https://doi.org/10.1161/ATVBAHA.116.307414

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    Intracellular Localization of Endogenous Mouse ABCG1 Is Mimicked by Both ABCG1-L550 and ABCG1-P550
    Elizabeth J. Tarling and Peter A. Edwards
    Arteriosclerosis, Thrombosis, and Vascular Biology. 2016;ATVBAHA.116.307414, originally published May 26, 2016
    https://doi.org/10.1161/ATVBAHA.116.307414
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