Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals

Elizabeth A. Jonas, John A. Hickman, Mushtaque Chachar, Brian M. Polster, Teresa A. Brandt, Yihru Fannjiang, Iva Ivanovska, Gorka Basañez, Kathleen W. Kinnally, Joshua Zimmerberg, J. Marie Hardwick, Leonard K. Kaczmarek

Research output: Contribution to journalArticle

Abstract

Neuronal death is often preceded by functional alterations at nerve terminals. Anti- and proapoptotic BCL-2 family proteins not only regulate the neuronal death pathway but also affect excitability of healthy neurons. We found that exposure of squid stellate ganglia to hypoxia, a death stimulus for neurons, causes a cysteine protease-dependent loss of full-length antiapoptotic BCL-xL, similar to previous findings in mammalian cells. Therefore, to determine the direct effect of the naturally occurring proapoptotic cleavage product of BCL-xL on mitochondria, recombinant N-truncated BCL-xL was applied to mitochondria inside the squid presynaptic terminal and to purified mitochondria isolated from yeast. N-truncated BCL-xL rapidly induced large multi-conductance channels with a maximal conductance significantly larger than those produced by full-length BCL-xL. This activity required the hydrophobic C terminus and the BH3 domain of BCL-xL. Moreover, N-truncated BCL-xL failed to produce any channel activity when applied to plasma membranes, suggesting that a component of the mitochondrial membrane is necessary for its actions. Consistent with this idea, the large channels induced by N-truncated BCL-xL are inhibited by NADH and require the presence of VDAC, a voltage-dependent anion channel present in the outer mitochondrial membrane. These observations suggest that the mitochondrial channels specific to full-length and N-truncated BCL-xL contribute to their opposite effects on synaptic transmission, and are consistent with their opposite effects on the cell death pathway.

Original languageEnglish (US)
Pages (from-to)13590-13595
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number37
DOIs
StatePublished - Sep 14 2004

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Presynaptic Terminals
Decapodiformes
Mitochondria
Mitochondrial Membranes
Voltage-Dependent Anion Channels
Stellate Ganglion
Neurons
Proteins
Cysteine Proteases
Synaptic Transmission
NAD
Cell Death
Yeasts
Cell Membrane

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals. / Jonas, Elizabeth A.; Hickman, John A.; Chachar, Mushtaque; Polster, Brian M.; Brandt, Teresa A.; Fannjiang, Yihru; Ivanovska, Iva; Basañez, Gorka; Kinnally, Kathleen W.; Zimmerberg, Joshua; Hardwick, J. Marie; Kaczmarek, Leonard K.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 101, No. 37, 14.09.2004, p. 13590-13595.

Research output: Contribution to journalArticle

Jonas, EA, Hickman, JA, Chachar, M, Polster, BM, Brandt, TA, Fannjiang, Y, Ivanovska, I, Basañez, G, Kinnally, KW, Zimmerberg, J, Hardwick, JM & Kaczmarek, LK 2004, 'Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals', Proceedings of the National Academy of Sciences of the United States of America, vol. 101, no. 37, pp. 13590-13595. https://doi.org/10.1073/pnas.0401372101
Jonas, Elizabeth A. ; Hickman, John A. ; Chachar, Mushtaque ; Polster, Brian M. ; Brandt, Teresa A. ; Fannjiang, Yihru ; Ivanovska, Iva ; Basañez, Gorka ; Kinnally, Kathleen W. ; Zimmerberg, Joshua ; Hardwick, J. Marie ; Kaczmarek, Leonard K. / Proapoptotic N-truncated BCL-xL protein activates endogenous mitochondrial channels in living synaptic terminals. In: Proceedings of the National Academy of Sciences of the United States of America. 2004 ; Vol. 101, No. 37. pp. 13590-13595.
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AU - Hickman, John A.

AU - Chachar, Mushtaque

AU - Polster, Brian M.

AU - Brandt, Teresa A.

AU - Fannjiang, Yihru

AU - Ivanovska, Iva

AU - Basañez, Gorka

AU - Kinnally, Kathleen W.

AU - Zimmerberg, Joshua

AU - Hardwick, J. Marie

AU - Kaczmarek, Leonard K.

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AB - Neuronal death is often preceded by functional alterations at nerve terminals. Anti- and proapoptotic BCL-2 family proteins not only regulate the neuronal death pathway but also affect excitability of healthy neurons. We found that exposure of squid stellate ganglia to hypoxia, a death stimulus for neurons, causes a cysteine protease-dependent loss of full-length antiapoptotic BCL-xL, similar to previous findings in mammalian cells. Therefore, to determine the direct effect of the naturally occurring proapoptotic cleavage product of BCL-xL on mitochondria, recombinant N-truncated BCL-xL was applied to mitochondria inside the squid presynaptic terminal and to purified mitochondria isolated from yeast. N-truncated BCL-xL rapidly induced large multi-conductance channels with a maximal conductance significantly larger than those produced by full-length BCL-xL. This activity required the hydrophobic C terminus and the BH3 domain of BCL-xL. Moreover, N-truncated BCL-xL failed to produce any channel activity when applied to plasma membranes, suggesting that a component of the mitochondrial membrane is necessary for its actions. Consistent with this idea, the large channels induced by N-truncated BCL-xL are inhibited by NADH and require the presence of VDAC, a voltage-dependent anion channel present in the outer mitochondrial membrane. These observations suggest that the mitochondrial channels specific to full-length and N-truncated BCL-xL contribute to their opposite effects on synaptic transmission, and are consistent with their opposite effects on the cell death pathway.

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