Proteomic mapping of ER-PM junctions identifies STIMATE as regulator of Ca2+ influx

Specialized junctional sites that connect the plasma membrane (PM) and endoplasmic reticulum (ER) play critical roles in controlling lipid metabolism and Ca2+ signaling1–4. Store operated Ca2+ entry mediated by dynamic STIM1-ORAI1 coupling represents a classical molecular event occurring at ER-PM junctions, but the protein composition and how previously-unrecognized protein regulators facilitate this process remain ill-defined. Using a combination of spatiallyUsers may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms CORRESPONDENCE: Y Zhou, Institute of Biosciences and Technology, Texas A&M University Health Science Center, 2121 W Holcombe Blvd, Houston, TX 77030, USA TEL +1-713-677-7483; [email protected] or to Y.W. ([email protected]). *These authors contributed equally to this work. AUTHOR CONTRIBUTIONS YZ and YW supervised and coordinated the study. LH, JJ, AQ and YZ designed and generated all the plasmid constructs. LH performed the BiFc assays. JJ, PT and LH generated the knockout cell lines. LH, YD, and MQD prepared the proteomic samples and performed the mass spectrometry analyses. GM, JJ, XL and YZ developed the in vitro assays, and carried out the experiments with assistance from LH, PT and YH. AS, JJ, YW and SLZ performed the Ca2+ influx assay. JJ, AS, AQ, LH, XZ, LC and YW performed all the fluorescence imaging and other cell-based experiments. XS contributed to the synthesis of biotin-phenol. YZ, JJ, YD, LH, AS, GM, YW and MQD analyzed data, with input from the other authors. PGH, YH and CLW provided intellectual inputs to the manuscript. JJ, YW and YZ wrote the manuscript. AUTHOR INFORMATION The authors state that they have no competing interests. Supplementary Information is available in the online version of the paper. HHS Public Access Author manuscript Nat Cell Biol. Author manuscript; available in PMC 2016 April 01. Published in final edited form as: Nat Cell Biol. 2015 October ; 17(10): 1339–1347. doi:10.1038/ncb3234. A uhor M anscript