An IRBIT homologue lacks binding activity to inositol 1,4,5-trisphosphate receptor due to the unique N-terminal appendage

Abstract

IRBIT is an inositol 1,4,5-trisphosphate (IP3) receptor (IP 3R)-binding protein that inhibits the activation of IP3R by competing with IP3 for the common binding site on IP3R. In this study, we characterize an IRBIT homologue, termed Long-IRBIT. Long-IRBIT is highly homologous to IRBIT (∼88%) and heteromerizes with IRBIT. In spite of complete conservation of critical amino acids required for the interaction with IP3R and comparable phosphorylations on critical four Ser residues for IP3R-binding, Long-IRBIT retains little ability to interact with IP3R. Deletion mutagenesis analysis revealed that this low affinity to IP3R is attributable to an inhibitory effect of the Long-IRBIT specific N-terminal appendage (LISN domain). Immunohistochemical analysis revealed the distinct distribution of Long-IRBIT and IRBIT in mouse cerebellar cortex, that is, Long-IRBIT is mainly expressed in interneurons such as basket cells, while IRBIT is mainly expressed in glial cells. Furthermore, Long-IRBIT, but not IRBIT, underwent phosphorylation during neuronal differentiation in neuroblastoma cells and this phosphorylation was dependent on the LISN domain. These results suggest that Long-IRBIT has a different function from IRBIT. © 2009 International Society for Neurochemistry.