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Yorodumi- EMDB-17691: 60-meric complex of dihydrolipoamide acetyltransferase (E2) of th... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-17691 | |||||||||
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Title | 60-meric complex of dihydrolipoamide acetyltransferase (E2) of the human pyruvate dehydrogenase complex (icosahedral symmetry) | |||||||||
Map data | ||||||||||
Sample |
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Keywords | pyruvate dehydrogenase complex / PDHc / E2 / cryo-EM / TRANSFERASE | |||||||||
Function / homology | Function and homology information dihydrolipoyllysine-residue acetyltransferase / dihydrolipoyllysine-residue acetyltransferase activity / acetyl-CoA biosynthetic process from pyruvate / : / pyruvate dehydrogenase complex / Glyoxylate metabolism and glycine degradation / Pyruvate metabolism / Regulation of pyruvate dehydrogenase (PDH) complex / Signaling by Retinoic Acid / tricarboxylic acid cycle ...dihydrolipoyllysine-residue acetyltransferase / dihydrolipoyllysine-residue acetyltransferase activity / acetyl-CoA biosynthetic process from pyruvate / : / pyruvate dehydrogenase complex / Glyoxylate metabolism and glycine degradation / Pyruvate metabolism / Regulation of pyruvate dehydrogenase (PDH) complex / Signaling by Retinoic Acid / tricarboxylic acid cycle / glucose metabolic process / mitochondrial matrix / intracellular membrane-bounded organelle / mitochondrion / identical protein binding Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.9 Å | |||||||||
Authors | Zdanowicz R / Afanasyev P / Boehringer D / Glockshuber R | |||||||||
Funding support | Switzerland, 1 items
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Citation | Journal: Sci Adv / Year: 2024 Title: Stoichiometry and architecture of the human pyruvate dehydrogenase complex. Authors: Rafal Zdanowicz / Pavel Afanasyev / Adam Pruška / Julian A Harrison / Christoph Giese / Daniel Boehringer / Alexander Leitner / Renato Zenobi / Rudi Glockshuber / Abstract: The pyruvate dehydrogenase complex (PDHc) is a key megaenzyme linking glycolysis with the citric acid cycle. In mammalian PDHc, dihydrolipoamide acetyltransferase (E2) and the dihydrolipoamide ...The pyruvate dehydrogenase complex (PDHc) is a key megaenzyme linking glycolysis with the citric acid cycle. In mammalian PDHc, dihydrolipoamide acetyltransferase (E2) and the dihydrolipoamide dehydrogenase-binding protein (E3BP) form a 60-subunit core that associates with the peripheral subunits pyruvate dehydrogenase (E1) and dihydrolipoamide dehydrogenase (E3). The structure and stoichiometry of the fully assembled, mammalian PDHc or its core remained elusive. Here, we demonstrate that the human PDHc core is formed by 48 E2 copies that bind 48 E1 heterotetramers and 12 E3BP copies that bind 12 E3 homodimers. Cryo-electron microscopy, together with native and cross-linking mass spectrometry, confirmed a core model in which 8 E2 homotrimers and 12 E2-E2-E3BP heterotrimers assemble into a pseudoicosahedral particle such that the 12 E3BP molecules form six E3BP-E3BP intertrimer interfaces distributed tetrahedrally within the 60-subunit core. The even distribution of E3 subunits in the peripheral shell of PDHc guarantees maximum enzymatic activity of the megaenzyme. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_17691.map.gz | 44 MB | EMDB map data format | |
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Header (meta data) | emd-17691-v30.xml emd-17691.xml | 17.2 KB 17.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_17691_fsc.xml | 15.2 KB | Display | FSC data file |
Images | emd_17691.png | 230.4 KB | ||
Filedesc metadata | emd-17691.cif.gz | 5.9 KB | ||
Others | emd_17691_half_map_1.map.gz emd_17691_half_map_2.map.gz | 337.7 MB 337.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-17691 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-17691 | HTTPS FTP |
-Validation report
Summary document | emd_17691_validation.pdf.gz | 774 KB | Display | EMDB validaton report |
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Full document | emd_17691_full_validation.pdf.gz | 773.6 KB | Display | |
Data in XML | emd_17691_validation.xml.gz | 24.7 KB | Display | |
Data in CIF | emd_17691_validation.cif.gz | 32.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17691 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-17691 | HTTPS FTP |
-Related structure data
Related structure data | 8piuMC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_17691.map.gz / Format: CCP4 / Size: 371.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Voxel size | X=Y=Z: 0.84 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #1
File | emd_17691_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_17691_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : 60-meric complex of dihydrolipoamide acetyltransferase (E2) of th...
Entire | Name: 60-meric complex of dihydrolipoamide acetyltransferase (E2) of the human pyruvate dehydrogenase complex |
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Components |
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-Supramolecule #1: 60-meric complex of dihydrolipoamide acetyltransferase (E2) of th...
Supramolecule | Name: 60-meric complex of dihydrolipoamide acetyltransferase (E2) of the human pyruvate dehydrogenase complex type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 3.6 MDa |
-Macromolecule #1: Dihydrolipoyllysine-residue acetyltransferase component of pyruva...
Macromolecule | Name: Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial type: protein_or_peptide / ID: 1 / Number of copies: 60 / Enantiomer: LEVO / EC number: dihydrolipoyllysine-residue acetyltransferase |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 59.811832 KDa |
Recombinant expression | Organism: Escherichia coli BL21(DE3) (bacteria) |
Sequence | String: MSLPPHQKVP LPSLSPTMQA GTIARWEKKE GDKINEGDLI AEVETDKATV GFESLEECYM AKILVAEGTR DVPIGAIICI TVGKPEDIE AFKNYTLDSS AAPTPQAAPA PTPAATASPP TPSAQAPGSS YPPHMQVLLP ALSPTMTMGT VQRWEKKVGE K LSEGDLLA ...String: MSLPPHQKVP LPSLSPTMQA GTIARWEKKE GDKINEGDLI AEVETDKATV GFESLEECYM AKILVAEGTR DVPIGAIICI TVGKPEDIE AFKNYTLDSS AAPTPQAAPA PTPAATASPP TPSAQAPGSS YPPHMQVLLP ALSPTMTMGT VQRWEKKVGE K LSEGDLLA EIETDKATIG FEVQEEGYLA KILVPEGTRD VPLGTPLCII VEKEADISAF ADYRPTEVTD LKPQVPPPTP PP VAAVPPT PQPLAPTPSA PCPATPAGPK GRVFVSPLAK KLAVEKGIDL TQVKGTGPDG RITKKDIDSF VPSKVAPAPA AVV PPTGPG MAPVPTGVFT DIPISNIRRV IAQRLMQSKQ TIPHYYLSID VNMGEVLLVR KELNKILEGR SKISVNDFII KASA LACLK VPEANSSWMD TVIRQNHVVD VSVAVSTPAG LITPIVFNAH IKGVETIAND VVSLATKARE GKLQPHEFQG GTFTI SNLG MFGIKNFSAI INPPQACILA IGASEDKLVP ADNEKGFDVA SMMSVTLSCD HRVVDGAVGA QWLAEFRKYL EKPITM LL UniProtKB: Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex, mitochondrial |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 80.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.8000000000000003 µm / Nominal defocus min: 1.6 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |