Purified TPC Isoforms Form NAADP Receptors with Distinct Roles for Ca²⁺ Signaling and Endolysosomal Trafficking

Intracellular Ca²⁺ signals constitute key elements in signal transduction. Of the three major Ca²⁺ mobilizing messengers described, the most potent, nicotinic acid adenine dinucleotide phosphate (NAADP) is the least well understood in terms of its molecular targets [1]. Recently, we showed that heterologous expression of two-pore channel (TPC) proteins enhances NAADP-induced Ca²⁺ release, whereas the NAADP response was abolished in pancreatic beta cells from Tpcn2 gene knockout mice [2]. However, whether TPCs constitute native NAADP receptors is unclear. Here we show that immunopurified endogenous TPC complexes possess the hallmark properties ascribed to NAADP receptors, including nanomolar ligand affinity [3-5]. Our study also reveals important functional differences between the three TPC isoforms. Thus, TPC1 and TPC2 both mediate NAADP-induced Ca²⁺ release, but the subsequent amplification of this trigger Ca²⁺ by IP₃Rs is more tightly coupled for TPC2. In contrast, TPC3 expression suppressed NAADP-induced Ca²⁺ release. Finally, increased TPC expression has dramatic and contrasting effects on endolysosomal structures and dynamics, implicating a role for NAADP in the regulation of vesicular trafficking. We propose that NAADP regulates endolysosomal Ca²⁺ storage and release via TPCs and coordinates endoplasmic reticulum Ca²⁺ release in a role that impacts on Ca²⁺ signaling in health and disease [6].