Lysosomal protease pathways to apoptosis: Cleavage of Bid, not pro-caspases, is the most likely route

Veronika Stoka, Boris Turk, Sharon L. Schendel, Tae Hyoung Kim, Tina Cirman, Scott J. Snipas, Lisa M. Ellerby, Dale Bredesen, Hudson Freeze, Magnus Abrahamson, Dieter Brömme, Stanislaw Krajewski, John C. Reed, Xiao-Ming Yin, Vito Turk, Guy S. Salvesen

Research output: Contribution to journalArticle

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Abstract

We investigated the mechanism of lysosome-mediated cell death using purified recombinant pro-apoptotic proteins, and cell-free extracts from the human neuronal progenitor cell line NT2. Potential effectors were either isolated lysosomes or purified lysosomal proteases. Purified lysosomal cathepsins B, H, K, L, S, and X or an extract of mouse lysosomes did not directly activate either recombinant caspase zymogens or caspase zymogens present in an NT2 cytosolic extract to any significant extent. In contrast, a cathepsin L-related protease from the protozoan parasite Trypanosoma cruzi, cruzipain, showed a measurable caspase activation rate. This demonstrated that members of the papain family can directly activate caspases but that mammalian lysosomal members of this family may have been negatively selected for caspase activation to prevent inappropriate induction of apoptosis. Given the lack of evidence for a direct role in caspase activation by lysosomal proteases, we hypothesized that an indirect mode of caspase activation may involve the Bcl-2 family member Bid. In support of this, Bid was cleaved in the presence of lysosomal extracts, at a site six residues down-stream from that seen for pathways involving capase 8. Incubation of mitochondria with Bid that had been cleaved by lysosomal extracts resulted in cytochrome c release. Thus, cleavage of Bid may represent a mechanism by which proteases that have leaked from the lysosomes can precipitate cytochrome c release and subsequent caspase activation. This is supported by the finding that cytosolic extracts from mice ablated in the bid gene are impaired in the ability to release cytochrome c in response to lysosome extracts. Together these data suggest that Bid represents a sensor that allows cells to initiate apoptosis in response to widespread adventitious proteolysis.

Original languageEnglish (US)
Pages (from-to)3149-3157
Number of pages9
JournalJournal of Biological Chemistry
Volume276
Issue number5
DOIs
StatePublished - Feb 2 2001
Externally publishedYes

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Caspases
Peptide Hydrolases
Apoptosis
Lysosomes
Chemical activation
Cytochromes c
Enzyme Precursors
Cathepsin H
Proteolysis
Cathepsin L
Cathepsin B
Apoptosis Regulatory Proteins
Mitochondria
Aptitude
Papain
Trypanosoma cruzi
Cell death
Cell Extracts
Recombinant Proteins
Precipitates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Stoka, V., Turk, B., Schendel, S. L., Kim, T. H., Cirman, T., Snipas, S. J., ... Salvesen, G. S. (2001). Lysosomal protease pathways to apoptosis: Cleavage of Bid, not pro-caspases, is the most likely route. Journal of Biological Chemistry, 276(5), 3149-3157. https://doi.org/10.1074/jbc.M008944200

Lysosomal protease pathways to apoptosis : Cleavage of Bid, not pro-caspases, is the most likely route. / Stoka, Veronika; Turk, Boris; Schendel, Sharon L.; Kim, Tae Hyoung; Cirman, Tina; Snipas, Scott J.; Ellerby, Lisa M.; Bredesen, Dale; Freeze, Hudson; Abrahamson, Magnus; Brömme, Dieter; Krajewski, Stanislaw; Reed, John C.; Yin, Xiao-Ming; Turk, Vito; Salvesen, Guy S.

In: Journal of Biological Chemistry, Vol. 276, No. 5, 02.02.2001, p. 3149-3157.

Research output: Contribution to journalArticle

Stoka, V, Turk, B, Schendel, SL, Kim, TH, Cirman, T, Snipas, SJ, Ellerby, LM, Bredesen, D, Freeze, H, Abrahamson, M, Brömme, D, Krajewski, S, Reed, JC, Yin, X-M, Turk, V & Salvesen, GS 2001, 'Lysosomal protease pathways to apoptosis: Cleavage of Bid, not pro-caspases, is the most likely route', Journal of Biological Chemistry, vol. 276, no. 5, pp. 3149-3157. https://doi.org/10.1074/jbc.M008944200
Stoka, Veronika ; Turk, Boris ; Schendel, Sharon L. ; Kim, Tae Hyoung ; Cirman, Tina ; Snipas, Scott J. ; Ellerby, Lisa M. ; Bredesen, Dale ; Freeze, Hudson ; Abrahamson, Magnus ; Brömme, Dieter ; Krajewski, Stanislaw ; Reed, John C. ; Yin, Xiao-Ming ; Turk, Vito ; Salvesen, Guy S. / Lysosomal protease pathways to apoptosis : Cleavage of Bid, not pro-caspases, is the most likely route. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 5. pp. 3149-3157.
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AU - Stoka, Veronika

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AU - Kim, Tae Hyoung

AU - Cirman, Tina

AU - Snipas, Scott J.

AU - Ellerby, Lisa M.

AU - Bredesen, Dale

AU - Freeze, Hudson

AU - Abrahamson, Magnus

AU - Brömme, Dieter

AU - Krajewski, Stanislaw

AU - Reed, John C.

AU - Yin, Xiao-Ming

AU - Turk, Vito

AU - Salvesen, Guy S.

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