The Role of the S4-S5 Linker and C-terminal Tail in Inositol 1,4,5-Trisphosphate Receptor Function

In previous studies we have suggested that spatial proximity of the C- and N-terminal domains of inositol 1,4,5-trisphosphate receptors (IP3Rs) may be critical for the channel gating mechanism. In the present study we have examined the sites of C-N interaction in more detail. We report that deletion mutations within the S4-S5 linker (amino acids 2418–2437) prevent co-immunoprecipitation of the C- and N-terminal domains, inhibit channel activity and enhance IP3 binding. We also show that a region of the C-terminal tail (amino acids 2694–2721), predicted to be a coiled-coil, is also required for channel activity. Circular dichroism spectroscopy and gel filtration studies confirm that this region has a helical structure with the ability to form tetramers. We propose a model in which IP3-induced conformational changes in the N-terminal domain are mechanically transmitted to the opening of the pore through an attachment to the S4-S5 linker. The coiled-coil domain in the C-terminal tail may play a critical role in maintaining the structural integrity of the channel.