The TRPM1 channel in ON-bipolar cells is gated by both the α and the βγ subunits of the G-protein Go

• Published in: Scientific reports 2016-02, Vol.6 (1), p.20940-20940, Article 20940
• Main Authors: Xu, Ying, Orlandi, Cesare, Cao, Yan, Yang, Shengyan, Choi, Chan-Il, Pagadala, Vijayakanth, Birnbaumer, Lutz, Martemyanov, Kirill A., Vardi, Noga
• Format: Article
• Language: English
• Subjects: 631/378/2613/1786; 631/443/376; Animals; Cell Line; Dark Adaptation; Darkness; Guanosine 5'-O-(3-Thiotriphosphate) - metabolism; Heterotrimeric GTP-Binding Proteins - genetics; Heterotrimeric GTP-Binding Proteins - metabolism; Humanities and Social Sciences; Humans; Ion Channel Gating; Light; Mice; Models, Animal; multidisciplinary; Mutation; Myristic Acid - pharmacology; Protein Binding; Rats; Retinal Bipolar Cells - metabolism; Retinal Bipolar Cells - radiation effects; Science; TRPM Cation Channels - metabolism
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep20940

Transmission from photoreceptors to ON bipolar cells in mammalian retina is mediated by a sign-inverting cascade. Upon binding glutamate, the metabotropic glutamate receptor mGluR6 activates the heterotrimeric G-protein Gα o β3γ13 and this leads to closure of the TRPM1 channel (melastatin). TRPM1 is thought to be constitutively open, but the mechanism that leads to its closure is unclear. We investigated this question in mouse rod bipolar cells by dialyzing reagents that modify the activity of either Gα o or Gβγ and then observing their effects on the basal holding current. After opening the TRPM1 channels with light, a constitutively active mutant of Gα o closed the channel, but wild-type Gα o did not. After closing the channels by dark adaptation, phosducin or inactive Gα o (both sequester Gβγ) opened the channel while the active mutant of Gα o did not. Co-immunoprecipitation showed that TRPM1 interacts with Gβ3 and with the active and inactive forms of Gα o . Furthermore, bioluminescent energy transfer assays indicated that while Gα o interacts with both the N- and the C- termini of TRPM1, Gβγ interacts only with the N-terminus. Our physiological and biochemical results suggest that both Gα o and Gβγ bind TRPM1 channels and cooperate to close them.