title: Differential mechanisms of action of the mucolipin synthetic agonist, ML-SA1, on insect TRPML and mammalian TRPML1.
abstract: Mucolipin synthetic agonist 1 (ML-SA1) was recently identified to activate mammalian TRPML channels and shown to alleviate lipid accumulation in lysosomes of cellular models of lysosome storage diseases, mucolipidosis type IV (MLIV) and Niemann-Pick's disease type C (NPC). Owning to its potential use in complimenting genetic studies in Drosophila melanogaster to elucidate the cellular and physiological functions of TRPML channels, we examined the effect of ML-SA1 on Drosophila TRPML expressed in HEK293 cells using whole-cell, inside-out, and whole-lysosome electrophysiological recordings. We previously showed that when expressed in HEK293 cells, Drosophila TRPML was localized and functional on both plasma membrane and endolysosome. We show here that in both inside-out patches excised from the plasma membrane and whole-lysosome recordings from enlarged endolysosome vacuoles, ML-SA1 failed to activate TRPML unless exogenous phosphatidylinositol 3,5-bisphosphate [PI(3,5)P2] was applied. At 1 μM ML-SA1, the sensitivity of TRPML to PI(3,5)P2 increased approximately by 10-fold and at 10 μM ML-SA1, the deactivation of PI(3,5)P2-evoked TRPML currents was markedly slowed. On the other hand, constitutive activation of TRPML by a mutation that mimics the varitint-waddler (Va) mutation of mouse TRPML3 rendered the insect channel sensitive to activation by ML-SA1 alone. Moreover, different from the insect TRPML, mouse TRPML1 was readily activated by ML-SA1 independent of PI(3,5)P2. Thus, our data reveal that while ML-SA1 acts as a true agonist at mouse TRPML1, it behaves as an allosteric activator of the Drosophila TRPML, showing dependence on and the ability to stabilize open conformation of the insect channels.
authors.affiliation: The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China; Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA.
authors.forename: Xinghua
authors.initials: X
authors.lastname: Feng
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Graduate Program in Cell and Regulatory Biology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiong
authors.affiliation: The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China; Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA.
authors.forename: Yungang
authors.initials: Y
authors.lastname: Lu
authors.affiliation: The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China; Center for Genomic Medicine, Houston Methodist Research Institute, Houston, TX 77030, USA. Electronic address: xuefengx@gmail.com.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Xia
authors.affiliation: Department of Integrative Biology and Pharmacology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA; Graduate Program in Cell and Regulatory Biology, The University of Texas Health Science Center at Houston, Houston, TX 77030, USA. Electronic address: michael.x.zhu@uth.tmc.edu.
authors.forename: Michael X
authors.initials: MX
authors.lastname: Zhu
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D029721:Drosophila Proteins / Q000187:drug effects* / Q000378:metabolism; D004331:Drosophila melanogaster; D057809:HEK293 Cells; D006801:Humans; D050356:Lipid Metabolism / Q000187:drug effects; D051379:Mice; D018408:Patch-Clamp Techniques; D018129:Phosphatidylinositol Phosphates / Q000494:pharmacology; D010797:Phthalimides / Q000494:pharmacology; D011804:Quinolines / Q000494:pharmacology; D013045:Species Specificity; D050051:Transient Receptor Potential Channels / Q000187:drug effects* / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; C416674:MCOLN1 protein, human; C000599935:ML-SA1 compound; C512321:Mcoln1 protein, mouse; D018129:Phosphatidylinositol Phosphates; D010797:Phthalimides; D011804:Quinolines; C578373:TRPML protein, Drosophila; D050051:Transient Receptor Potential Channels; C106336:phosphatidylinositol 3,5-diphosphate
keywords: Allosteric activator; Calcium; Lysosome storage disease; Neurodegeneration; TRP channels; TRPML