Fragmented Inositol 1,4,5-Trisphosphate Receptors Retain Tetrameric Architecture and Form Functional Ca2+ Release Channels

• Published in: The Journal of biological chemistry 2013-04, Vol.288 (16), p.11122-11134
• Main Authors: Alzayady, Kamil J., Chandrasekhar, Rahul, Yule, David I.
• Format: Article
• Language: English
• Subjects: Animals; Apoptosis; Apoptosis - drug effects; Apoptosis - physiology; Calcium Channels - genetics; Calcium Channels - metabolism; Calcium Intracellular Release; Calcium Signaling; Caspase; Enzyme Inhibitors - pharmacology; Inositol 1,4,5-Trisphosphate; Inositol 1,4,5-Trisphosphate Receptors; Inositol 1,4,5-Trisphosphate Receptors - genetics; Inositol 1,4,5-Trisphosphate Receptors - metabolism; Mice; Protein Multimerization - drug effects; Protein Multimerization - physiology; Protein Structure, Quaternary; Proteolysis; Rats; Signal Transduction; Staurosporine - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M113.453241

Inositol 1,4,5-trisphosphate receptor isoforms are a family of ubiquitously expressed ligand-gated channels encoded by three individual genes. The proteins are localized to membranes of intracellular Ca2+ stores and play pivotal roles in Ca2+ homeostasis. Previous studies have demonstrated that IP3R1 is cleaved by the intracellular proteases calpain and caspase both in vivo and in vitro. However, the resultant cleavage products are poorly defined, and the functional consequences of these proteolytic events are not fully understood. We demonstrate that IP3R1 is cleaved during staurosporine-induced apoptosis, yielding N-terminal fragments encompassing the ligand-binding domain and the majority of the central modulatory domain together with a C-terminal fragment containing the channel domain and cytosolic tail. Notably, these fragments remain associated with the membrane after initiation of apoptotic cleavage. Furthermore, when recombinant IP3R1 fragments, corresponding to those predicted to be generated by caspase or calpain cleavage, are stably coexpressed in cells, they physically associate and form functional channels. These data provide novel insights regarding the regulation of IP3R1 during proteolysis and provide direct evidence that polypeptide continuity is not required for IP3R activation and Ca2+ release. Background: Proteolytic cleavage and disruption of inositol 1,4,5-trisphosphate receptor (IP3R) architecture may contribute to the initiation/progression of apoptosis. Results: Proteolytic cleavage products of the IP3R remain membrane-associated, and these fragments form functional tetrameric channels. Conclusion: Fragmentation of the IP3R does not inevitably lead to loss of function. Significance: Peptide continuity is not required for IP3R function, and IP3R activation may persist after protease cleavage during apoptosis.