Key resources table
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| MCU | Cell Signaling | Cat#D2Z3B; RRID: AB_2721812 |
| EMRE | Santa Cruz | Cat#86337; RRID: AB_2250685 |
| MICU1 | Atlas Antibodies | Cat#HPA034780; RRID: AB_10696934 |
| MICU2 | Abcam | Cat#ab101465 |
| MICU3 | Atlas Antibodies | Cat# HPA024771; RRID: AB_1848022 |
| VDAC | Abcam | Cat# ab14734 RRID: AB_443084 |
| COX1 | Abcam | Cat#ab14705; RRID: AB_2084810 |
| COX2 | Abcam | Cat#ab14734 |
| Tim23 | Santa Cruz | Cat#sc-5114463 |
| FLAG | Sigma-Aldrich | Cat#F1804; RRID: AB_262044 |
| YME1L1 | Proteintech | Cat#11510-1-AP; RRID: AB_2217459 |
| AFG3L2 | Proteintech | Cat#14631-1-AP; RRID: AB_2242420 |
| Letm1 | Abcam | Cat#ab55434; RRID: AB_943985 |
| β-actin | Santa Cruz | Cat#sc-69879; RRID: AB_1119529 |
| NDUFS1 | Santa Cruz | Cat#sc-271510; RRID: AB10655669 |
| NDUFS3 | Santa Cruz | Cat#sc-374282; RRID: AB_10991315 |
| SDHA | Santa Cruz | Cat#sc-390381 |
| Rieske | Santa Cruz | Cat#sc-271609 RRID: AB_10655682 |
| ATP5F1 | Santa Cruz | Cat#sc-514419 |
| 1D4 | Home made | N/A |
| PDH-E1ɑ | Santa Cruz | Cat#sc-377092 RRID: AB_2716767 |
| S293 phosphorylated PDH-E1ɑ | Millipore | Cat#ABS204 RRID: AB_11205754 |
| IRDye 680RD goat-anti rabbit | LI-COR | Cat#925-68071; RRID: AB_2721181 |
| IRDye 800CW goat-anti mouse | LI-COR | Cat#925-32210; RRID: AB_2687825 |
| IRDye 680RD goat-anti mouse | LI-COR | Cat#925-68070 RRID: AB_2651128 |
| Alkaline phosphatase conjugated anti-rabbit secondary antibody | Promega | Cat# S373B |
| Bacterial and virus strains | ||
| E. coli: DH5ɑ competent cells | New England Biolabs | Cat#C2987H |
| E. coli: BL21(DE3) | Christopher Miller | Tsai et al., 2012 |
| Biological samples | ||
| Left ventricular tissues of non-failing human hearts | Adult Cardiac Tissue Bank at the University of Colorado Denver-Anschutz Medical campus | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Lipofectamine 3000 | Life Technologies | Cat#L3000150 |
| Lipofectamine 2000 | Life Technologies | Cat#11668019 |
| DDM, anagrade | Anatrace | Cat#D310 |
| RIPA buffer | Santa Cruz | Cat#sc-24948 |
| Intercept blocking buffer (TBS-based) | LI-COR | Cat#927-60001 |
| PVDF membranes | LI-COR | Cat#926-31097 |
| Protease inhibitor cocktail tablet | Roche | Cat#05892970001 |
| TALON cobalt affinity resin | Takara | Cat#635504 |
| CNBr-activated Sepharose 4B | GE Healthcare | Cat#17043001 |
| Cycloheximide | Sigma | Cat#C7698 |
| Oligomycin | Cayman | Cat#11342 |
| Rotenone | Cayman | Cat#13995 |
| Antimycin A | Sigma | Cat#A8674 |
| Ionomycin | Cayman | Cat#11932 |
| Carbachol | Cayman | Cat#114486 |
| 45Ca2+ radioisotope | Perkin-Elmer | Cat#NEZ013001MC |
| 35S-methionine | Perkin-Elmer | Cat#NEG009A001MC |
| Cysteine/methionine free medium | Sigma | Cat#D0422 |
| Protein A beads | Santa Cruz | Cat#sc-2001 |
| Thapsigargin | Cayman | Cat#10522 |
| Digitonin | Sigma-Aldrich | Cat#D141 |
| Calcium green 5N | Life Technologies | Cat#C3737 |
| TMRM | Invitrogen | Cat#T668 |
| FCCP | Cayman | Cat#15218 |
| Doxorubicin | Tokyo Chemical Industry | Cat#D4193 |
| Cisplatin | Tokyo Chemical Industry | Cat#D3371 |
| Annexin V CF488A | Biotium | Cat#29083 |
| Propidium Iodide | Cayman | Cat#14289 |
| Hoescht 33258 | Cayman | Cat#16756 |
| Luciferin | ThermoFisher | Cat#PI88291 |
| Critical commercial assays | ||
| Seahorse XF Real-time ATP Rate Assay Kit | Agilent | Cat#103591-100 |
| QuickChange II Kit | Agilent | Cat#200524 |
| BCA assay kit | Thermo | Cat#23225 |
| NativePAGE Sample Prep kit | Invitrogen | Cat#BN2008 |
| NativePAGE Running Buffer kit | Invitrogen | Cat#BN2007 |
| Deposited data | ||
| Unprocessed images used in the figures | This paper | doi:10.17632/shrtrdfhyt.1 |
| Experimental models: Cell lines | ||
| HEK 293T: wild type | ATCC | Cat#CRL-11268; RRID: CVCL_1926 |
| HeLa: wild type | ATCC | Cat#CCL-2; RRID: CVCL_0030 |
| U2OS: wild type | Cell Technologies Resource Center in the University of Colorado Anschutz Medical Campus | https://medschool.cuanschutz.edu/colorado-cancer-center/research/shared-resources/cell-technologies/cell-culture-services |
| HEK 293T: MICU1-KO | Home made | Tsai et al., 2017 |
| HEK 293T: MICU2-KO | Home made | Fan et al., 2020 |
| HEK 293T: MICU1-MICU2-KO | Home made | Fan et al., 2020 |
| HEK 239T: C463R-MICU1 (CRISPR base edited) | This paper | N/A |
| MEF cells: wild type | Vivek Garg | Garg et al., 2021 |
| MEF cells: MICU1-KO | Vivek Garg | Garg et al., 2021 |
| MEF cells: MICU2-KO | Vivek Garg | Garg et al., 2021 |
| Experimental models: Organisms/strains | ||
| C57BL/6J mouse: wild type | Jackson Laboratory | Cat#000664; RRID: IMSR_JAX:000664 |
| Sprague Dawley rats: wild type | Charles River Laboratory | Cat#001; RRID: RGD_734476 |
| Oligonucleotides | ||
| sgRNA sequence to mutate native MICU1 to C463R in HEK cells: 5’-ACACCTTCCTGTGCACATTTCCACAG | This paper | N/A |
| Recombinant DNA | ||
| MCU, EMRE, MICU1 or MICU2 in pcDNA3.1(+) | Home made | Tsai et al., 2016; Tasi et al., 2017 |
| MICU1 or MICU2 in pLJM1 | This paper | N/A |
| pLKO.hygro constructs to knockdown YME1L1 or AFG3L2 | Home made | Tasi et al., 2017 |
| pCMV_ABEmax | Koblan et al., 2018 | Addgene Cat#112095 |
| MLM3636 sgRNA expression plasmid | Keith Joung | Addgene Cat#43860 |
| Mito-GCaMP6f | Timothy Ryan | Ashrafi et al., 2020 |
| Cyto-GCaMP6f | Chen et al., 2013 | Addgene Cat#40755 |
| Mito-Sypher | Poburko et al., 2011 | Addgene Cat#48251 |
| Cyto-Sypher | Poburko et al., 2011 | Addgene Cat#48250 |
| Mito-roGFP2-Orp1 | Gutscher et al., 2009 | Addgene Cat#64992 |
| Mitochondria-targeted luciferase | Paolo Pinton | Morciano et al., 2017 |
| Mito-GEM-GECO1 | Zhao et al., 2011 | Addgene Cat#32461 |
| MBP-MICU1 in pET24a | This paper | N/A |
| MICU2 in pETDuet | This paper | N/A |
| Software and algorithms | ||
| Igor pro (version 8) | WaveMetrics | https://www.wavemetrics.com/products/igorpro |
| Image Studio (version 5.2) | LI-COR | https://www.licor.com/bio/image-studio/ |
| ImageJ (Mac OS X) | ImageJ website | https://imageJ.nih.gov/ij |
| Time Series Analyzer plug-in (version 3) | Balaji J | https://imagej.nih.gov/ij/plugins/time-series.html |