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Calpeptin, not calpain, directly inhibits an ion channel of the inner mitochondrial membrane

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Abstract

The permeability transition pore (PTP) of inner mitochondrial membranes is a large conductance pathway for ions up to 1500 Da which opening is responsible for ion equilibration and loss of membrane potential in apoptosis and thus in several neurodegenerative diseases. The PTP can be regulated by the Ca2+-activated mitochondrial K channel (BK). Calpains are Ca2+-activated cystein proteases; calpeptin is an inhibitor of calpains. We wondered whether calpain or calpeptin can modulate activity of PTP or BK. Patch clamp experiments were performed on mitoplasts of rat liver (PTP) and of an astrocytoma cell line (BK). Channel-independent open probability (P o) was determined (PTP) and, taking into account the number of open levels, NPo by single channel analysis (BK). We find that PTP in the presence of Ca2+ (200 μM) is uninfluenced by calpain (13 nM) and shows insignificant decrease by the calpain inhibitor calpeptin (1 μM). The NPo of the BK is insensitive to calpain (54 nM), too. However, it is significantly and reversibly inhibited by the calpain inhibitor calpeptin (IC50 = 42 μM). The results agree with calpeptin-induced activation of the PTP via inhibition of the BK. Screening experiments with respirometry show calpeptin effects, fitting to inhibition of the BK by calpeptin, and strong inhibition of state 3 respiration.

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Acknowledgments

We are indebted to F. N. Gellerich and Z. Gizatullina for advice and helpful discussions throughout the respirometry experiments. Mrs. K. Kaiser, C. Höhne, and J. Witzke gave us technical support, and Mrs. D. Koch from Leibniz Institute of Neurobiology, Magdeburg, kindly provided us with rat livers. Financial support by Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE) is gratefully acknowledged.

Conflicts of interest

None of the authors has any conflicts of interest.

(Detlef Siemen on behalf of the authors)

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Authors and Affiliations

  1. Department of Neurology, Otto-von-Guericke-Universität, Leipziger Str. 44, D-39120, Magdeburg, Germany

    Maria Derksen & Detlef Siemen

  2. Ophthalmology, SRH Zentralklinikum Suhl, D-98527, Suhl, Germany

    Christian Vorwerk

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  1. Maria Derksen
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  2. Christian Vorwerk
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  3. Detlef Siemen
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Correspondence to Detlef Siemen.

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Handling Editor: Reimer Stick

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Derksen, M., Vorwerk, C. & Siemen, D. Calpeptin, not calpain, directly inhibits an ion channel of the inner mitochondrial membrane. Protoplasma 253, 835–843 (2016). https://doi.org/10.1007/s00709-015-0846-x

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  • DOI: https://doi.org/10.1007/s00709-015-0846-x

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