TWO PORE CHANNELS RECEPTORS INTERACTING PROTEINS STUDY: AN AUTOPHAGIC ROLE

Abstract

Autophagy is an evolutionarily conserved lysosomal degradation pathway, yet the underlying mechanisms remain poorly understood. In previous studies, we showed the Nicotinic acid adenine dinucleotide phosphate (NAADP) as a potent Ca2+ mobilizing messenger in astrocytes, since NAADP releases Ca2+ from the endo-lysosomal system and this effect was mediated by the two-pore channels (TPCs), especially TPC type 2. Some independent studies have been shown that the specificity of NAADP-mediated Ca2+ release is inhibited by Ned-19. But, whether and how TPCs and NAADP act in astrocytes is unclear and the functions on autophagy pathway have yet to be defined. To better understand how TPCs functions are regulated, a proteomic screening by Mass Spectrometric of TPC1 and TPC2 interacting proteins will be performed. Because TPCs might be NAADP-sensitive Ca2+-permeable channel in lysosomes, will be examined the NAADP-AM on interacting proteins regulation in TPC1 or 2-overexpressing cells. Furthermore, the levels of interacting proteins autophagy-related also will be studied under NAADP or under autophagic inducers effects through western blotting and immunoprecipitation experiments. Taken together, our results further will demonstrate that the activation of NAADP/TPCs signaling might compromise the differently the autophagy progression, suggesting the potential physiological of NAADP/TPCs in central nervous system.