Neurotoxicity induces cleavage of p35 to p25 by calpain

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Abstract

Cyclin-dependent kinase 5 (cdk5) and its neuron-specific activator p35 are required for neurite outgrowth and cortical lamination1,2,3. Proteolytic cleavage of p35 produces p25, which accumulates in the brains of patients with Alzheimer's disease4. Conversion of p35 to p25 causes prolonged activation and mislocalization of cdk5. Consequently, the p25/cdk5 kinase hyperphosphorylates tau, disrupts the cytoskeleton and promotes the death (apoptosis) of primary neurons. Here we describe the mechanism of conversion of p35 to p25. In cultured primary cortical neurons, excitotoxins, hypoxic stress and calcium influx induce the production of p25. In fresh brain lysates, addition of calcium can stimulate cleavage of p35 to p25. Specific inhibitors of calpain, a calcium-dependent cysteine protease, effectively inhibit the calcium-induced cleavage of p35. In vitro, calpain directly cleaves p35 to release a fragment with relative molecular mass 25,000. The sequence of the calpain cleavage product corresponds precisely to that of p25. Application of the amyloid β-peptide Aβ(1–42) induces the conversion of p35 to p25 in primary cortical neurons. Furthermore, inhibition of cdk5 or calpain activity reduces cell death in Aβ-treated cortical neurons. These observations indicate that cleavage of p35 to p25 by calpain may be involved in the pathogenesis of Alzheimer's disease.

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Figure 1: Neurotoxicity induces cleavage of p35 to p25.
Figure 2: Ca2+ is necessary for p35 cleavage in vitro.
Figure 3: Calpain directly cleaves p35 to p25.
Figure 4: Conversion of p35 to p25 in primary cortical neurons is mediated by calpain and requires Ca2+.
Figure 5: Aβ(1–42) induces conversion of p35 to p25; inhibition of cdk5 or calpain reduces Aβ(1–42)-induced cell death.

References

  1. 1

    Ohshima, T. et al. Targeted disruption of the cyclin-dependent kinase 5 gene results in abnormal corticogenesis, neuronal pathology and perinatal death. Proc. Natl Acad. Sci. USA 93, 11173– 11178 (1996).

  2. 2

    Chae, T. et al. Mice lacking p35, a neuronal specific activator of Cdk5, display cortical lamination defects, seizures, and adult lethality. Neuron 18, 29–42 ( 1997).

  3. 3

    Nikolic, M., Dudek, H., Kwon, Y. T., Ramos, Y. F. & Tsai, L. H. The cdk5/p35 kinase is essential for neurite outgrowth during neuronal differentiation. Genes Dev. 10, 816–825 (1996).

  4. 4

    Patrick, G. N. et al. Conversion of p35 to p25 de-regulates cdk5 activity and promotes neurodegeneration. Nature 402, 615– 622 (1999).

  5. 5

    Wang, K. K. W. in Calpain: Pharmacology and Toxicology of Calcium-Dependent Protease (eds Wang, K. K. W. & Yuen, P.-W.) 77–102 (Taylor & Francis, Philadelphia, 1999).

  6. 6

    Sorimachi, H., Ishiura, S. & Suzuki, K. Structure and physiological function of calpains. Biochem. J. 328, 721–732 (1997).

  7. 7

    Lew, J. et al. A brain-specific activator of cyclin-dependent kinase 5. Nature 371, 423–426 ( 1994).

  8. 8

    Yankner, B. A. Mechanisms of neuronal degeneration in Alzheimer's disease. Neuron 16, 921–932 ( 1996).

  9. 9

    Alvarez, A., Toro, R., Caceres, A. & Maccioni, R. B. Inhibition of tau phosphorylating protein kinase cdk5 prevents β-amyloid-induced neuronal death. FEBS Lett. 459, 421– 426 (1999).

  10. 10

    Arispe, N., Pollard, H. B. & Rojas, E. Giant multilevel cation channels formed by Alzheimer disease amyloid beta protein [A beta P-(1-40)] in bilayer membranes. Proc. Natl Acad. Sci. USA 90, 10573– 10577 (1993).

  11. 11

    Hartmann, H., Eckert, A. & Muller, W. E. β-Amyloid protein amplifies calcium signalling in central neurons from the adult mouse. Biochem. Biophys. Res. Commun. 194, 1216–1220 ( 1993).

  12. 12

    Mattson, M. P. et al. β-Amyloid peptides destabilize calcium homeostasis and render human cortical neurons vulnerable to excitotoxicity. J. Neurosci. 12, 376–389 ( 1992).

  13. 13

    Pascale, A. & Etcheberrigaray, R. Calcium alterations in Alzheimer's disease: pathophysiology, models and therapeutic opportunities. Pharmacol. Res. 39, 81–88 (1999).

  14. 14

    Grynspan, R., Griffin, W. R., Cataldo, A., Katayama, S. & Nixon, R. A. Active site-directed antibodies identify calpain II as an early-appearing and pervasive component of neurofibrillary pathology in Alzheimer's disease. Brain Res. 763, 145–158 (1997).

  15. 15

    Nixon, R. A. et al. Calcium-activated neutral proteinase (calpain) system in aging and Alzheimer's disease. Ann. NY Acad. Sci. 747, 77–91 (1994).

  16. 16

    Saito, K., Elce, J. S., Hamos, J. E. & Nixon, R. A. Widespread activation of calcium-activated neutral proteinase (calpain) in the brain in Alzheimer disease: a potential molecular basis for neuronal degeneration. Proc. Natl Acad. Sci. USA 90, 2628– 2632 (1993).

  17. 17

    Iwamoto, N., Thangnipon, W., Crawford, C. & Emson, P. C. Localization of calpain immunoreactivity in senile plaques and in neurones undergoing neurofibrillary degeneration in Alzheimer's disease. Brain Res. 561, 177–180 (1991).

  18. 18

    Selkoe, D. J. Translating cell biology into therapeutic advances in Alzheimer's disease. Nature 399, A23–A31 (1999).

  19. 19

    Seisjo, B. K. in Brain Energy Metabolism 454 (Wiley, New York, 1978).

  20. 20

    Nikolic, M., Chou, M. M., Lu, W., Mayer, B. J. & Tsai, L. H. The p35/Cdk5 kinase is a neuron-specific Rac effector that inhibits Pak1 activity. Nature 395, 194–198 (1998).

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Acknowledgements

We thank J. Elce for the m-calpain antibody; A. Bonni and V. Tannoch for reading this manuscript; M. Ahlijanian, B. Yankner, E. Wang, K. Kosik and members of the Tsai lab for discussions. This work was partially supported by NIH grants to L.-H.T. L.-H.T. is an assistant investigator of the Howard Hughes Medical Institute, a Rita Allen Foundation scholar and a recipient of an Ester A. and Joseph Kligenstein Fund grant.

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Correspondence to Li-Huei Tsai.

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