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Calpain-mediated cleavage negatively regulates the expression level of ABCG1

  • Author Footnotes
    1 These authors contributed equally to this work.
    Natsuki Hori
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Laboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.
    Hisamitsu Hayashi
    Correspondence
    Corresponding author at: Laboratory of Molecular Pharmacokinetics, Department of Medical Pharmaceutics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Tel.: +81 3 5841 4773; fax: +81 3 5841 4766.
    Footnotes
    1 These authors contributed equally to this work.
    Affiliations
    Laboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
    Search for articles by this author
  • Yuichi Sugiyama
    Correspondence
    Corresponding author at: Laboratory of Molecular Pharmacokinetics, Department of Medical Pharmaceutics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. Tel.: +81 3 5841 4770; fax: +81 3 5841 4766.
    Affiliations
    Laboratory of Molecular Pharmacokinetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan
    Search for articles by this author
  • Author Footnotes
    1 These authors contributed equally to this work.

      Abstract

      Objective

      The ATP-binding cassette transporter ABCG1 mediates cholesterol efflux from macrophages, and prevents the progression of macrophage foam-cell formation. Much less is known about the regulatory mechanism of ABCG1, although its physiological importance is becoming clearer. Here, we show the role of calpain in ABCG1 degradation.

      Methods and results

      Purified μ-calpain cleaved ABCG1 in crude membrane fractions prepared from human ABCG1-expressing HEK293 (ABCG1-HEK) cells. In ABCG1-HEK cells, calpeptin treatment, a calpain inhibitor, inhibited ABCG1 degradation, and thereby increased the expression and cholesterol efflux function of ABCG1. Biotinylation study demonstrated greater ABCG1 induction with calpeptin treatment in cell surface than that in whole cell lysates. Together with the result that increased ABCG1 expression with calpeptin treatment was observed under clathrin heavy-chain (CHC) knockdown conditions, where ABCG1 internalization was prevented, calpain is considered to catalyze ABCG1 cleavage on the plasma membrane. In mouse peritoneal macrophages as well as in ABCG1-HEK cells, calpeptin treatment inhibited ABCG1 degradation and enhanced ABCG1 expression, even under CHC-depleted conditions.

      Conclusion

      These observations indicate that calpain promotes ABCG1 degradation by cleaving cell surface-resident ABCG1, and consequently reduces the expression and cholesterol efflux function of ABCG1. Inhibition of ABCG1 cleavage by calpain could be a novel approach to suppress the progression of atherosclerosis.

      Keywords

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