Neurotoxicity induces cleavage of p35 to p25 by calpain

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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.


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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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