Calpain-cleaved Type 1 Inositol 1,4,5-Trisphosphate Receptor (InsP3R1) Has InsP3-independent Gating and Disrupts Intracellular Ca2+ Homeostasis
| Autor: | Catherine M. Kopil, Horia Vais, Robert W. Neumar, J. Kevin Foskett, Don On Daniel Mak, King-Ho Cheung, Adam P. Siebert |
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| Rok vydání: | 2011 |
| Předmět: |
Male
endocrine system Endoplasmic Reticulum Biochemistry Cell Line Purkinje Cells chemistry.chemical_compound Calpain - metabolism - pharmacology Animals Homeostasis Inositol 1 4 5-Trisphosphate Receptors Rats Long-Evans Inositol Calcium Signaling Molecular Biology Ion channel Calcium signaling Cell Death biology Voltage-dependent calcium channel Calpain Ionomycin Endoplasmic reticulum Calcium - metabolism Inositol 1 4 5-Trisphosphate Receptors - metabolism Molecular Bases of Disease Cell Biology Calcium Signaling - drug effects Rats Cell biology carbohydrates (lipids) Calcium Ionophores chemistry Proteolysis biology.protein Calcium Calcium Channels Ion Channel Gating - drug effects Ion Channel Gating Intracellular |
| Zdroj: | Journal of Biological Chemistry. 286:35998-36010 |
| ISSN: | 0021-9258 |
| Popis: | The type 1 inositol 1,4,5-trisphosphate receptor (InsP(3)R1) is a ubiquitous intracellular Ca(2+) release channel that is vital to intracellular Ca(2+) signaling. InsP(3)R1 is a proteolytic target of calpain, which cleaves the channel to form a 95-kDa carboxyl-terminal fragment that includes the transmembrane domains, which contain the ion pore. However, the functional consequences of calpain proteolysis on channel behavior and Ca(2+) homeostasis are unknown. In the present study we have identified a unique calpain cleavage site in InsP(3)R1 and utilized a recombinant truncated form of the channel (capn-InsP(3)R1) corresponding to the stable, carboxyl-terminal fragment to examine the functional consequences of channel proteolysis. Single-channel recordings of capn-InsP(3)R1 revealed InsP(3)-independent gating and high open probability (P(o)) under optimal cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) conditions. However, some [Ca(2+)](i) regulation of the cleaved channel remained, with a lower P(o) in suboptimal and inhibitory [Ca(2+)](i). Expression of capn-InsP(3)R1 in N2a cells reduced the Ca(2+) content of ionomycin-releasable intracellular stores and decreased endoplasmic reticulum Ca(2+) loading compared with control cells expressing full-length InsP(3)R1. Using a cleavage-specific antibody, we identified calpain-cleaved InsP(3)R1 in selectively vulnerable cerebellar Purkinje neurons after in vivo cardiac arrest. These findings indicate that calpain proteolysis of InsP(3)R1 generates a dysregulated channel that disrupts cellular Ca(2+) homeostasis. Furthermore, our results demonstrate that calpain cleaves InsP(3)R1 in a clinically relevant injury model, suggesting that Ca(2+) leak through the proteolyzed channel may act as a feed-forward mechanism to enhance cell death. link_to_OA_fulltext |
| Databáze: | OpenAIRE |
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