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Lysosomal chymotrypsin B potentiates apoptosis via cleavage of Bid

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Abstract

We have reported that chymotrypsin B (CtrB) is not just a digestive enzyme but is also stored in lysosomes. Herein, we demonstrated a broad distribution of CtrB and explored the involvement of CtrB in apoptosis. Exposure of RH-35 cells to H2O2 or palmitate induced the redistribution of lysosomal CtrB into the cytoplasm as a result of lysosomal membrane permeabilization (LMP). Suppression of CtrB significantly blocked apoptosis, while overexpression of CtrB sensitized apoptosis markedly. CtrB could cleave Bid under neutral conditions. In RH-35 cells with Bid silenced, apoptosis induced by CtrB protein was attenuated, suggesting that CtrB mediates apoptosis of RH-35 cells mainly through processing Bid. Our data also suggest that LMP occurs earlier than mitochondrial outer membrane permeabilization; Bid activation initiated by caspase-8 might be reinforced by CtrB in consequence of LMP, which causes a positive feedback loop leading to the accumulation of tBid, and results in lysosome- and mitochondrion-dependent apoptosis.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (Grants 30900246 and 30770492), the National Basic Research Program of China (Grants 2006CB705600, 2006CB911001 and 2010CB833700), and the K. C. Wong Education Foundation, Hong Kong. We would like to thank Dr. L. Gráf (Eötvös Lorárd University, Hungary) for kindly providing plasmid encoding rat chymotrypsin B, and Prof. R. S. Chen (Institute of Biophysics, CAS) and Prof. Frank Dowd (Creighton University, USA) for valuable suggestions and discussions. The authors are also indebted to W. M. Zhong, W. Li, H. J. Zhou and P. T. Jiang for their technical assistance.

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Correspondence to Taotao Wei or Fuyu Yang.

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K. Zhao and X. Zhao contributed equally to this work.

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Zhao, K., Zhao, X., Tu, Y. et al. Lysosomal chymotrypsin B potentiates apoptosis via cleavage of Bid. Cell. Mol. Life Sci. 67, 2665–2678 (2010). https://doi.org/10.1007/s00018-010-0356-0

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