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Demonstration of calmodulin-sensitive calcium translocation by isolated osteoclast plasma membrane vesicles

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Summary

Plasma membrane vesicles were prepared from chicken osteoclasts, and active calcium transport was demonstrated in a spectrofluorimetric assay using the fluorescent calcium concentration indicator, fura-2. Transport activity was inhibited by quercetin (10 μM), sodium vanadate (10 μM), and the anticalmodulin agents, compound 48/80 (20 and 200 μg/ml) and calmidazolium (10 and 20 μM). The transport rate (Vmax, 1.3 nmol/mg protein/min) was not altered in the presence of the protonophore, nigericin (1 μM), indicating that proton transport was not driving calcium transport. Release of accumulated calcium in the vesicles occurred with the addition of bromo-A23187 (5 μM) or ionomycin (5 μM). Increasing calcium transport occurred with increasing calcium concentration. Finally, the calmodulin content of the vesicles was demonstrated to be 54–134 U/mg protein. These results demonstrate that a calmodulin-sensitive, ATP-dependent calcium transporter is present in the osteoclast plasma membrane.

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Authors and Affiliations

  1. Department of Molecular and Cell Biology, The Pennsylvania State University, 468A North Frear, 16802, University Park, Pennsylvania, USA

    Petrus J. Bekker & Carol V. Gay

  2. Department of Poultry Science, The Pennsylvania State University, 468A North Frear, 16802, University Park, Pennsylvania, USA

    Carol V. Gay

Authors
  1. Petrus J. Bekker
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  2. Carol V. Gay
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Bekker, P.J., Gay, C.V. Demonstration of calmodulin-sensitive calcium translocation by isolated osteoclast plasma membrane vesicles. Calcif Tissue Int 51, 312–316 (1992). https://doi.org/10.1007/BF00334493

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  • DOI: https://doi.org/10.1007/BF00334493

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