Single-Particle CryoEM of mammalian V-ATPase with the TLDc domain protein mEAK7 bound (Various Datasets)

REL 2021-11-08 2022-09-05 2022-09-05 Single-Particle CryoEM of mammalian V-ATPase with the TLDc domain protein mEAK7 bound (Various Datasets) 0000-0001-6656-6320 Yong Zi Tan The Hospital for Sick Children Research Institute 686 Bay Street Toronto Ontario Canada M5G 0A4 0000-0003-0566-2209 John Leonard Rubinstein The Hospital for Sick Children Research Institute/University of Toronto 686 Bay Street Toronto Ontario Canada M5G 0A4 Tan YZ Abbas YM Wu JZ Wu D Keon KA Hesketh GG Bueler SA Gingras AC Robinson CV Grinstein S Rubinstein JL 12.7 10.6019/EMPIAR-10874 EMDB molecule EMD-26385 EMD-26386 EMD-26387 EMD-26388 Tan YZ Abbas YM Wu JZ Wu D Keon KA Hesketh GG Bueler SA Gingras AC Robinson CV Grinstein S Rubinstein JL CryoEM of endogenous mammalian V-ATPase interacting with the TLDc protein mEAK-7 Life science alliance Life Sci Alliance 11 5 2022 English 10.26508/lsa.202201527 35794005

The movies, micrographs and particle stacks of pig kidney V-ATPase with various constructs of mEAK-7, under various biochemical and imaging conditions.

Tan YZ Abbas YM Wu JZ Wu D Hesketh GG Bueler SA Gingras AC Robinson CV Grinstein S Rubinstein JL CryoEM of endogenous mammalian V-ATPase interacting with the TLDc protein mEAK-7 bioRxiv 2021 10.1101/2021.11.03.466369

The movies, micrographs and particle stacks of pig kidney V-ATPase with various constructs of mEAK-7, under various biochemical and imaging conditions.

Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra /data/01_VATPase-mEAK-7_Tundra/Movies micrographs - multiframe MRCS MRCS 1957 24 1 24 32 BIT FLOAT 4096 0.7512 4096 0.7512

Movies collected using Tundra. 24 frames were used, pixel size is 0.7512 Å/pixel, total dose of 40 e-/Å2. Compressed using pbzip2.

data/01_VATPase-mEAK-7_Tundra/Micrographs data/01_VATPase-mEAK-7_Tundra/Particles/State*.star data/01_VATPase-mEAK-7_Tundra/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra /data/01_VATPase-mEAK-7_Tundra/Micrographs micrographs - single frame MRC MRC 1957 1 32 BIT FLOAT 4096 0.7512 4096 0.7512

Micrographs collected using Tundra. Aligned and dose weighted using UCSF MotionCor2.

data/01_VATPase-mEAK-7_Tundra/Micrographs data/01_VATPase-mEAK-7_Tundra/Particles/State*.star data/01_VATPase-mEAK-7_Tundra/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7 collected using Tundra /data/01_VATPase-mEAK-7_Tundra/Particles picked particles - single frame - processed MRCS MRCS 1 19158 1 19158 32 BIT FLOAT 320 1.461 320 1.461

Final particle stack after particle polishing using Relion.

data/01_VATPase-mEAK-7_Tundra/Micrographs data/01_VATPase-mEAK-7_Tundra/Particles/State*.star data/01_VATPase-mEAK-7_Tundra/Particles Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 /data/02_VATPase-mEAK-7_Krios/Movies micrographs - multiframe MRCS MRCS 5142 30 1 29 32 BIT FLOAT 4096 1.3225 4096 1.3225

Movies collected using Titan Krios and Falcon 4. 30 frames were used, pixel size is 1.3225 Å/pixel, total dose of 36 e-/Å2. Compressed using pbzip2. Last frame should be discarded during processing.

data/02_VATPase-mEAK-7_Krios/Micrographs data/02_VATPase-mEAK-7_Krios/Particles/VATPAseMEAK7-Polished.star data/02_VATPase-mEAK-7_Krios/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 /data/02_VATPase-mEAK-7_Krios/Micrographs micrographs - single frame MRC MRC 5142 1 32 BIT FLOAT 4096 1.3225 4096 1.3225

Micrographs collected using Titan Krios and Falcon 4. Aligned and dose weighted using UCSF MotionCor2.

data/02_VATPase-mEAK-7_Krios/Micrographs data/02_VATPase-mEAK-7_Krios/Particles/VATPAseMEAK7-Polished.star data/02_VATPase-mEAK-7_Krios/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7 collected using Titan Krios and Falcon4 /data/02_VATPase-mEAK-7_Krios/Particles picked particles - multiframe - processed MRCS MRCS 1 186338 1 186338 32 BIT FLOAT 360 1.3225 360 1.3225

Final particle stack after particle polishing using Relion.

data/02_VATPase-mEAK-7_Krios/Micrographs data/02_VATPase-mEAK-7_Krios/Particles/VATPAseMEAK7-Polished.star data/02_VATPase-mEAK-7_Krios/Particles Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 /data/03_VATPase-mEAK-7deltaCterm_Krios/Movies micrographs - multiframe MRCS MRCS 2013 30 1 29 32 BIT FLOAT 4096 1.3225 4096 1.3225

Movies collected using Titan Krios and Falcon 4. 30 frames were used, pixel size is 1.3225 Å/pixel, total dose of 37 e-/Å2. Compressed using pbzip2. Last frame should be discarded during processing.

data/03_VATPase-mEAK-7deltaCterm_Krios/Micrographs data/03_VATPase-mEAK-7deltaCterm_Krios/Particles/VATPase-mEAK7NtermTLDc_Polish.star data/03_VATPase-mEAK-7deltaCterm_Krios/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 /data/03_VATPase-mEAK-7deltaCterm_Krios/Micrographs micrographs - single frame MRC MRC 2013 1 32 BIT FLOAT 4096 1.3225 4096 1.3225

Micrographs collected using Titan Krios and Falcon 4. Aligned and dose weighted using UCSF MotionCor2.

data/03_VATPase-mEAK-7deltaCterm_Krios/Micrographs data/03_VATPase-mEAK-7deltaCterm_Krios/Particles/VATPase-mEAK7NtermTLDc_Polish.star data/03_VATPase-mEAK-7deltaCterm_Krios/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7deltaCterm collected using Titan Krios and Falcon4 /data/03_VATPase-mEAK-7deltaCterm_Krios/Particles picked particles - single frame - processed MRCS MRCS 1 107515 1 107515 32 BIT FLOAT 4096 1.3225 4096 1.3225

Final particle stack after particle polishing using Relion.

data/03_VATPase-mEAK-7deltaCterm_Krios/Micrographs data/03_VATPase-mEAK-7deltaCterm_Krios/Particles/VATPase-mEAK7NtermTLDc_Polish.star data/03_VATPase-mEAK-7deltaCterm_Krios/Particles Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 /data/04_VATPase-mEAK-7_ATP_Krios/Movies micrographs - multiframe MRCS MRCS 5498 30 1 29 32 BIT FLOAT 4096 1.3225 4096 1.3225

Movies collected using Titan Krios and Falcon 4. 30 frames were used, pixel size is 1.3225 Å/pixel, total dose of 44 e-/Å2. Compressed using pbzip2. Last frame should be discarded during processing.

data/04_VATPase-mEAK-7_ATP_Krios/Micrographs data/04_VATPase-mEAK-7_ATP_Krios/Particles/VATPAseMEAK7-10mMATP-Polished.star data/04_VATPase-mEAK-7_ATP_Krios/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 /data/04_VATPase-mEAK-7_ATP_Krios/Micrographs micrographs - single frame MRC MRC 5498 1 32 BIT FLOAT 4096 1.3225 4096 1.3225

Micrographs collected using Titan Krios and Falcon 4. Aligned and dose weighted using UCSF MotionCor2.

data/04_VATPase-mEAK-7_ATP_Krios/Micrographs data/04_VATPase-mEAK-7_ATP_Krios/Particles/VATPAseMEAK7-10mMATP-Polished.star data/04_VATPase-mEAK-7_ATP_Krios/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with ATP collected using Titan Krios and Falcon4 /data/04_VATPase-mEAK-7_ATP_Krios/Particles picked particles - single frame - processed MRCS MRCS 1 253892 1 253892 32 BIT FLOAT 4096 1.3225 4096 1.3225

Final particle stack after particle polishing using Relion.

data/04_VATPase-mEAK-7_ATP_Krios/Micrographs data/04_VATPase-mEAK-7_ATP_Krios/Particles/VATPAseMEAK7-10mMATP-Polished.star data/04_VATPase-mEAK-7_ATP_Krios/Particles Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 /data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Movies micrographs - multiframe MRCS MRCS 2468 30 1 29 32 BIT FLOAT 4096 1.3225 4096 1.3225

Movies collected using Titan Krios and Falcon 4. 30 frames were used, pixel size is 1.3225 Å/pixel, total dose of 41 e-/Å2. Compressed using pbzip2. Last frame should be discarded during processing.

data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Micrographs data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles/VATPAseMEAK7-EGTAEDTA-Polished.star data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 /data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Micrographs micrographs - single frame MRC MRC 2468 1 32 BIT FLOAT 4096 1.3225 4096 1.3225

Micrographs collected using Titan Krios and Falcon 4. Aligned and dose weighted using UCSF MotionCor2.

data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Micrographs data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles/VATPAseMEAK7-EGTAEDTA-Polished.star data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with EDTA/EGTA collected using Titan Krios and Falcon4 /data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles picked particles - single frame - processed MRCS MRCS 1 102191 1 102191 32 BIT FLOAT 360 1.3225 360 1.3225

Final particle stack after particle polishing using Relion.

data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Micrographs data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles/VATPAseMEAK7-EGTAEDTA-Polished.star data/05_VATPase-mEAK-7_EDTA-EGTA_Krios/Particles Unaligned multiframe movies of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 /data/06_VATPase-mEAK-7_Calcium_Glacios/Movies micrographs - multiframe EER EER 3029 1 32 BIT FLOAT 4096 0.895 4096 0.895

Movies collected using Glacios with Selectris X and Falcon 4. EER format was used, pixel size is 0.895 Å/pixel, total dose of 40 e-/Å2.

data/06_VATPase-mEAK-7_Calcium_Glacios/Micrographs data/06_VATPase-mEAK-7_Calcium_Glacios/Particles/VATPaseMEAK7-20mMCalcium_PolishRelion_320.star data/06_VATPase-mEAK-7_Calcium_Glacios/Particles Aligned and dose-weighted micrographs of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 /data/06_VATPase-mEAK-7_Calcium_Glacios/Micrographs micrographs - single frame MRC MRC 3029 1 32 BIT FLOAT 4096 0.895 4096 0.895

Micrographs collected using Titan Krios and Falcon 4. Aligned and dose weighted using UCSF MotionCor2.

data/06_VATPase-mEAK-7_Calcium_Glacios/Micrographs data/06_VATPase-mEAK-7_Calcium_Glacios/Particles/VATPaseMEAK7-20mMCalcium_PolishRelion_320.star data/06_VATPase-mEAK-7_Calcium_Glacios/Particles Polished particles of Pig Kidney V-ATPase bound to mEAK-7 with Calcium collected using Glacios with Selectris X and Falcon 4 /data/06_VATPase-mEAK-7_Calcium_Glacios/Particles picked particles - single frame - processed MRCS MRCS 1 123373 1 123373 32 BIT FLOAT 320 1.56625 320 1.56625

Final particle stack after particle polishing using Relion.

data/06_VATPase-mEAK-7_Calcium_Glacios/Micrographs data/06_VATPase-mEAK-7_Calcium_Glacios/Particles/VATPaseMEAK7-20mMCalcium_PolishRelion_320.star data/06_VATPase-mEAK-7_Calcium_Glacios/Particles