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Characterization of G Protein-coupled Signal Transduction of the Drosophila GPCR Methuselah
Zhang Jing1,2, Zhang Ru1, Ye Chenli1,2, Xie Xin1,2*
1Shanghai Key Laboratory of Signaling and Disease Research, Laboratory of Receptor-based Bio-medicine, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China; 2State Key Laboratory of Drug Research, the Nat
Abstract: Drosophila Methuselah (MTH) is a member of the G protein-coupled receptor (GPCR) super family. Mutation of MTH receptor leads to extended life span and resistance to various forms of stress in fruit fly. However， the signal transduction of MTH at cellular level has rarely been studied. Using HEK293 cell line which stably expresses MTH， we investigated its G protein-coupling preference. Firstly， we confirmed the expression and biological activity of MTH in HEK293 cells by using immunofluorescent staining， Western blot analysis and calcium mobilization assay. N-stunted， the endogenous ligand of MTH， induced calcium mobilization in cells expressing MTH. The calcium signal was insensitive to PTX pretreatment， indicating MTH is probably coupling to Gq/11 rather than Gi/o pathway. Then we discovered that activation of MTH did not induce any change in the intracellular cAMP level in HEK293 cells， suggesting that MTH is not coulpled to Gs or Gi/o. Finally， we showed that activationof MTH could induce MAP kinase ERK1/2 phosphorylation. Our results revealed major signal transduction pathways of MTH which might facilitate further research of this receptor and help us understanding its biological functions.
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