Calpain inhibitor causes accumulation of ubiquitinated P-glycoprotein at the cell surface: possible role of calpain in P-glycoprotein turnover.

  • Authors:
    • K Ohkawa
    • T Asakura
    • K Takada
    • T Sawai
    • Y Hashizume
    • Y Okawa
    • N Yanaihara
  • View Affiliations

  • Published online on: October 1, 1999     https://doi.org/10.3892/ijo.15.4.677
  • Pages: 677-763
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Abstract

P-glycoprotein (Pgp) is a plasma-membrane glycoprotein that confers multi-drug resistance (MDR) on cells and displays ATP-driven drug pumping. The possible contribution of calpain-mediated proteolytic pathways to the functional regulation of the Pgp molecule was evaluated using K562/DXR, MDR cells. N-Acetyl-L-leucyl-L-leucyl-norleucinal was effluxed by Pgp, but N-benzyloxycarbonyl-L-leucyl-L-leucinal (zLLal), an inhibitor of calpain, retarded the degradation of Pgp leading to accumulation of the molecule largely at the cell surface membrane. Treatment with brefeldin A did not obstruct the zLLal-induced Pgp accumulation. NH4Cl increased the cytoplasmic Pgp level, with a slight to significant decrease at the cell surface membrane. Ubiquitin-ELISA and western blot analysis confirmed that the Pgp molecule, which accumulated mainly at the cell surface, was ubiquitinated. However, lactacystin did not show any accumulation of Pgp in either the cytoplasm or the cell surface membrane, suggesting that the proteasome did not participate in the phenomenon. Additionally, the Pgp was limitedly proteolyzed by calpain into two 98 kDa and 69 kDa, fragments within one minute. Despite the increased accumulation of Pgp at the cell surface after treatment with calpain inhibitor, the cytoplasmic doxorubicin level of the cells treated with a calpain inhibitor was higher than that of non-treated cells and approached that of parental cells. These results indicated that calpain involved Pgp turnover and that calpain inhibition induced ubiquitinated Pgp-accumulation mainly at the cell surface membrane with a reduction in its own functions suggesting that the modulation of Pgp-turnover involves MDR-reversal by another approach.

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Oct 1999
Volume 15 Issue 4

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Ohkawa K, Asakura T, Takada K, Sawai T, Hashizume Y, Okawa Y and Yanaihara N: Calpain inhibitor causes accumulation of ubiquitinated P-glycoprotein at the cell surface: possible role of calpain in P-glycoprotein turnover.. Int J Oncol 15: 677-763, 1999.
APA
Ohkawa, K., Asakura, T., Takada, K., Sawai, T., Hashizume, Y., Okawa, Y., & Yanaihara, N. (1999). Calpain inhibitor causes accumulation of ubiquitinated P-glycoprotein at the cell surface: possible role of calpain in P-glycoprotein turnover.. International Journal of Oncology, 15, 677-763. https://doi.org/10.3892/ijo.15.4.677
MLA
Ohkawa, K., Asakura, T., Takada, K., Sawai, T., Hashizume, Y., Okawa, Y., Yanaihara, N."Calpain inhibitor causes accumulation of ubiquitinated P-glycoprotein at the cell surface: possible role of calpain in P-glycoprotein turnover.". International Journal of Oncology 15.4 (1999): 677-763.
Chicago
Ohkawa, K., Asakura, T., Takada, K., Sawai, T., Hashizume, Y., Okawa, Y., Yanaihara, N."Calpain inhibitor causes accumulation of ubiquitinated P-glycoprotein at the cell surface: possible role of calpain in P-glycoprotein turnover.". International Journal of Oncology 15, no. 4 (1999): 677-763. https://doi.org/10.3892/ijo.15.4.677