Abstract
Altering intracellular calcium levels is known to partially restore mutant enzyme homeostasis in several lysosomal storage diseases, but why? We hypothesized that endoplasmic reticulum (ER) calcium increases enhance the folding, trafficking and function of these mutant misfolding- and degradation-prone lysosomal enzymes by increasing chaperone function. Here we report that increasing ER calcium levels by reducing ER calcium efflux through the ryanodine receptor, using antagonists or RNAi, or by promoting ER calcium influx by SERCA2b overexpression enhances mutant glucocerebrosidase (GC) homeostasis in cells derived from individuals with Gaucher's disease. Post-translational regulation of the calnexin folding pathway by an elevated ER calcium concentration seems to enhance the capacity of this chaperone system to fold mutant misfolding-prone enzymes, increasing the folded mutant GC population that can engage the trafficking receptor at the expense of ER-associated degradation, increasing the lysosomal GC concentration and activity.
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Change history
14 May 2010
In the version of this article initially published online, three errors of sense were introduced into the text. Also, Figure 5b was incorrectly cited where Figure 5c should have been cited and an additional gene was erroneously listed as a chaperone not changing after diltiazem or dantrolene treatment in reference to Supplementary Figure 9a. These errors have been corrected for the print, PDF and HTML versions of this article.
21 May 2010
In the version of this article initially published, in the main text one of the cell lines used was erroneously given the mutant designation L444P in reference to Figure 6a and another cell line did not include this designation in reference to Supplementary Figure 11a,b. The errors have been corrected in the HTML and PDF versions of the article.
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Acknowledgements
We thank the following for their generosity in providing us with plasmids: W.E. Balch (The Scripps Research Institute) for pcDNA3.1+VSVG, E. Beutler (The Scripps Research Institute) for WT GC cDNA, M. Brenner (Harvard Medical School) for Apr-M8-CNX, K. Green (University of California, Irvine) for SERCA2 cDNA and T. Mizushima (Kumamoto University) for pCR(HA) and pcDNA3.1-GRP78. We thank H. Aerts (University of Amsterdam) for the mouse monoclonal anti-GC 8E4, W.E. Balch (The Scripps Research Institute) for mouse monoclonal anti-VSVG, M. Fukuda (Burnham Institute) for the rabbit anti-LAMP2 and C. Fearns for critical feedback on the manuscript. This work was supported by the NIH (DK75295), the Skaggs Institute for Chemical Biology and the Lita Annenberg Hazen Foundation. A.E.P. was supported by the NIH (GM084027).
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D.S.T.O. performed the majority of the experiments, analyzed the data and wrote the initial draft of the paper. T.-W.M. performed the initial RyR inhibitor experiments and all the Ca2+ measurement experiments and collaborated on the analysis of the data. The ER Ca2+ levels were measured in the A.E.P. laboratory. J.W.K. supervised the work and managed the final publication.
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J.W.K. is a cofounder, shareholder and paid consultant for Proteostasis Therapeutics, Inc., and although this company is not now pursuing any lysosomal storage diseases, that could happen in the future.
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Supplementary Figures 1–13, Supplementary Table 1 and Supplementary Methods (PDF 593 kb)
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Ong, D., Mu, TW., Palmer, A. et al. Endoplasmic reticulum Ca2+ increases enhance mutant glucocerebrosidase proteostasis. Nat Chem Biol 6, 424–432 (2010). https://doi.org/10.1038/nchembio.368
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DOI: https://doi.org/10.1038/nchembio.368
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