Structure of the EF-hand domain of polycystin-2 suggests a mechanism for Ca~(2+) -dependent regulation of polycystin-2 channel activity

摘要

The C-terminal cytoplasmic tail of polycystin-2 (PC2/TRPP2), a Ca~(2+) -permeable channel, is frequently mutated or truncated in autosomal dominant polycystic kidney disease. We have previously shown that this tail consists of three functional regions: an EF-hand domain (PC2-EF, 720-797), a flexible linker (798-827), and an oligomeric coiled coil domain (828-895). We found that PC2-EF binds Ca~(2+) at a single site and undergoes Ca~(2+) -dependent conformational changes, suggesting it is an essential element of Ca~(2+) -sensitive regulation of PC2 activity. Here we describe the NMR structure and dynamics of Ca~(2+) -bound PC2-EF. Human PC2-EF contains a divergent non-Ca~(2+) -binding helix-loop-helix (HLH) motif packed against a canonical Ca~(2+) -binding EF-hand motif. This HLH motif may have evolved from a canonical EF-hand found in invertebrate PC2 homologs. Temperature-dependent steady-state NOE experiments and NMR R_1 and R_2 relaxation rates correlate with increased molecular motion in the EF-hand, possibly due to exchange between apo and Ca~(2+) -bound states, consistent with a role for PC2-EF as a Ca~(2+) -sensitive regulator. Structure-based sequence conservation analysis reveals a conserved hydrophobic surface in the same region, which may mediate Ca~(2+) -dependent protein interactions. We propose that Ca~(2+)-sensing by PC2-EF is responsible for the cooperative nature of PC2 channel activation and inhibition. Based on our results, we present a mechanism of regulation of the Ca~(2+) dependence of PC2 channel activity by PC2-EF.