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MICU1 Interacts with the D-Ring of the MCU Pore to Control Its Ca Flux and Sensitivity to Ru360.

Mol. Cell. 2018; 
Paillard Melanie,Csordás György,Huang Kai-Ting,Várnai Peter,Joseph Suresh K,Hajnóczky Gy�
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Recombinant Antibody Services Molecular Cell 72, 778–785, November 15, 2018 785 STAR+METHODS KEY RESOURCES TABLE REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit anti-MICU1 Sigma-Aldrich HPA037480; RRID: AB_10696934 Rabbit anti-MCU Sigma-Aldrich HPA016480; RRID: AB_2071893 Rabbit anti-MICU2 Abcam ab101465; RRID: AB_10711219 Mouse anti-mtHsp70 Thermo Fisher MA3-028; RRID: AB_325474 Rabbit anti-prohibitin Abcam ab28172; RRID: AB_777457 Anti-HA polyclonal Invitrogen 71-5500; RRID: AB_2533988 Anti-FLAG M2 Sigma-Aldrich F1804; RRID: AB_262044 Anti-Myc monoclonal Precision AG10004 Chemicals, Peptides, and Recombinant Proteins Anti-DYKDDDDK (FLAG) beads Genscript A00187-200 Ruthenium Red Sigma R2751 Fura-2 (AM) Teflabs 0-103 Fura-2 (salt) Teflabs 0-104 Fura-2 low affinity (AM) Teflabs 0-136 Fura-2 low affinity (salt) Teflabs 0-137 Thapsigargin Enzo Life Sciences BML-PE180-0005 CGP-37157 Enzo Life Sciences BML-CM119-0005 Lipofectamine 3000 Life Technologies L3000008 Complete protease inhibitor cocktail (EDTA free) Roche 11873580001 Protein A Sepharose Abcam ab193256 Critical Commercial Assays DC Protein Assay Biorad 5000112 Experimental Models: Cell Lines MICU1loxp/loxp MEF Antony et al. Get A Quote

摘要

Proper control of the mitochondrial Ca uniporter's pore (MCU) is required to allow Ca-dependent activation of oxidative metabolism and to avoid mitochondrial Ca overload and cell death. The MCU's gatekeeping and cooperative activation is mediated by the Ca-sensing MICU1 protein, which has been proposed to form dimeric complexes anchored to the EMRE scaffold of MCU. We unexpectedly find that MICU1 suppresses inhibition of MCU by ruthenium red/Ru360, which bind to MCU's DIME motif, the selectivity filter. This led us to recognize in MICU1's sequence a putative DIME interacting domain (DID), which is required for both gatekeeping and cooperative activation of MCU and for cell survival. Thus, we propose ... More

关键词

Ca(2+) uptake,calcium signaling,cooperativity,mitochondria,oxidative stress,thres