+Open data
-Basic information
Entry | Database: PDB / ID: 8e2k | |||||||||
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Title | Cryo-EM structure of BIRC6/HtrA2-S306A | |||||||||
Components |
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Keywords | LIGASE / Ubiquitin / E3 ligase / Apoptosis / Autophagy / IAP | |||||||||
Function / homology | Function and homology information HtrA2 peptidase / pentacyclic triterpenoid metabolic process / negative regulation of mitophagy in response to mitochondrial depolarization / spongiotrophoblast layer development / positive regulation of cysteine-type endopeptidase activity involved in apoptotic signaling pathway / regulation of autophagy of mitochondrion / CD40 receptor complex / ceramide metabolic process / : / labyrinthine layer development ...HtrA2 peptidase / pentacyclic triterpenoid metabolic process / negative regulation of mitophagy in response to mitochondrial depolarization / spongiotrophoblast layer development / positive regulation of cysteine-type endopeptidase activity involved in apoptotic signaling pathway / regulation of autophagy of mitochondrion / CD40 receptor complex / ceramide metabolic process / : / labyrinthine layer development / programmed cell death / ALK mutants bind TKIs / positive regulation of extrinsic apoptotic signaling pathway in absence of ligand / serine-type endopeptidase complex / Flemming body / adult walking behavior / response to herbicide / microtubule organizing center / execution phase of apoptosis / positive regulation of protein targeting to mitochondrion / cysteine-type endopeptidase inhibitor activity / ubiquitin conjugating enzyme activity / positive regulation of cysteine-type endopeptidase activity involved in apoptotic process / protein autoprocessing / Signaling by ALK fusions and activated point mutants / regulation of multicellular organism growth / negative regulation of cell cycle / neuron development / cellular response to interferon-beta / negative regulation of oxidative stress-induced intrinsic apoptotic signaling pathway / forebrain development / cellular response to retinoic acid / mitochondrion organization / serine-type peptidase activity / regulation of cytokinesis / mitochondrial membrane / negative regulation of extrinsic apoptotic signaling pathway / protein catabolic process / RING-type E3 ubiquitin transferase / trans-Golgi network / mitochondrial intermembrane space / cytoplasmic side of plasma membrane / cellular response to growth factor stimulus / spindle pole / ubiquitin-protein transferase activity / intrinsic apoptotic signaling pathway in response to DNA damage / unfolded protein binding / cellular response to heat / cellular response to oxidative stress / regulation of cell population proliferation / peptidase activity / midbody / neuron apoptotic process / negative regulation of neuron apoptotic process / cell population proliferation / protein ubiquitination / cytoskeleton / endosome / cell cycle / positive regulation of apoptotic process / cell division / protein phosphorylation / serine-type endopeptidase activity / centrosome / apoptotic process / positive regulation of cell population proliferation / chromatin / endoplasmic reticulum membrane / negative regulation of apoptotic process / endoplasmic reticulum / mitochondrion / proteolysis / membrane / identical protein binding / nucleus / cytosol Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.21 Å | |||||||||
Authors | Hunkeler, M. / Fischer, E.S. | |||||||||
Funding support | United States, 2items
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Citation | Journal: Science / Year: 2023 Title: Structures of BIRC6-client complexes provide a mechanism of SMAC-mediated release of caspases. Authors: Moritz Hunkeler / Cyrus Y Jin / Eric S Fischer / Abstract: Tight regulation of apoptosis is essential for metazoan development and prevents diseases such as cancer and neurodegeneration. Caspase activation is central to apoptosis, and inhibitor of apoptosis ...Tight regulation of apoptosis is essential for metazoan development and prevents diseases such as cancer and neurodegeneration. Caspase activation is central to apoptosis, and inhibitor of apoptosis proteins (IAPs) are the principal actors that restrain caspase activity and are therefore attractive therapeutic targets. IAPs, in turn, are regulated by mitochondria-derived proapoptotic factors such as SMAC and HTRA2. Through a series of cryo-electron microscopy structures of full-length human baculoviral IAP repeat-containing protein 6 (BIRC6) bound to SMAC, caspases, and HTRA2, we provide a molecular understanding for BIRC6-mediated caspase inhibition and its release by SMAC. The architecture of BIRC6, together with near-irreversible binding of SMAC, elucidates how the IAP inhibitor SMAC can effectively control a processive ubiquitin ligase to respond to apoptotic stimuli. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8e2k.cif.gz | 2.2 MB | Display | PDBx/mmCIF format |
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PDB format | pdb8e2k.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8e2k.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/e2/8e2k ftp://data.pdbj.org/pub/pdb/validation_reports/e2/8e2k | HTTPS FTP |
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-Related structure data
Related structure data | 27841MC 8e2dC 8e2eC 8e2fC 8e2gC 8e2hC 8e2iC 8e2jC C: citing same article (ref.) M: map data used to model this data |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 535317.188 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: BIRC6, KIAA1289 / Production host: Homo sapiens (human) / Strain (production host): Expi293 / References: UniProt: Q9NR09, Ligases #2: Protein | Mass: 35961.887 Da / Num. of mol.: 3 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: HTRA2, OMI, PRSS25 / Production host: Escherichia coli (E. coli) / Strain (production host): LOBSTR / References: UniProt: O43464, HtrA2 peptidase |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Baculoviral IAP repeat-containing protein 6 / Type: COMPLEX / Entity ID: all / Source: MULTIPLE SOURCES | ||||||||||||||||||||
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Molecular weight | Value: 1.174 MDa / Experimental value: NO | ||||||||||||||||||||
Source (natural) | Organism: Homo sapiens (human) | ||||||||||||||||||||
Source (recombinant) | Organism: Homo sapiens (human) / Strain: Expi293 | ||||||||||||||||||||
Buffer solution | pH: 7.4 | ||||||||||||||||||||
Buffer component |
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Specimen | Conc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES / Details: added CHAPSO to 0.4 mM | ||||||||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | ||||||||||||||||||||
Vitrification | Instrument: LEICA EM GP / Cryogen name: ETHANE / Humidity: 90 % / Chamber temperature: 283.15 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS Details: Data collection in counting mode, using multi-shot scheme (9 holes per stage position, 3 movies per hole) |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELDBright-field microscopy / Nominal magnification: 105000 X / Nominal defocus max: 2100 nm / Nominal defocus min: 800 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Average exposure time: 2.5 sec. / Electron dose: 60.659 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 15309 |
EM imaging optics | Energyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV |
Image scans | Width: 5760 / Height: 4092 / Movie frames/image: 50 / Used frames/image: 1-50 |
-Processing
Software | Name: PHENIX / Version: 1.20.1_4487: / Classification: refinement | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Particle selection | Num. of particles selected: 2049607 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
3D reconstruction | Resolution: 3.21 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 73712 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Atomic model building | Protocol: OTHER / Space: REAL | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refine LS restraints |
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