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Yorodumi- PDB-8u2c: Gaussian mixture model based single particle refinement - ABC tra... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8u2c | ||||||
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Title | Gaussian mixture model based single particle refinement - ABC transporter (inhibitor-bound ABCG2 from EMPIAR-10374) | ||||||
Components |
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Keywords | MEMBRANE PROTEIN / ABC transporter | ||||||
Function / homology | Function and homology information biotin transmembrane transporter activity / biotin transport / riboflavin transport / riboflavin transmembrane transporter activity / renal urate salt excretion / sphingolipid transporter activity / urate metabolic process / urate transmembrane transporter activity / Abacavir transmembrane transport / organic anion transport ...biotin transmembrane transporter activity / biotin transport / riboflavin transport / riboflavin transmembrane transporter activity / renal urate salt excretion / sphingolipid transporter activity / urate metabolic process / urate transmembrane transporter activity / Abacavir transmembrane transport / organic anion transport / external side of apical plasma membrane / sphingolipid biosynthetic process / organic anion transmembrane transporter activity / Sphingolipid de novo biosynthesis / xenobiotic transport across blood-brain barrier / export across plasma membrane / ABC-type xenobiotic transporter / Paracetamol ADME / transepithelial transport / Ciprofloxacin ADME / cellular detoxification / ABC-type xenobiotic transporter activity / NFE2L2 regulating MDR associated enzymes / Heme biosynthesis / Heme degradation / efflux transmembrane transporter activity / xenobiotic transmembrane transporter activity / transport across blood-brain barrier / ATPase-coupled transmembrane transporter activity / mitochondrial membrane / brush border membrane / Iron uptake and transport / transmembrane transport / membrane raft / apical plasma membrane / protein homodimerization activity / ATP hydrolysis activity / nucleoplasm / ATP binding / identical protein binding / plasma membrane Similarity search - Function | ||||||
Biological species | Homo sapiens (human) Mus musculus (house mouse) | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 2.5 Å | ||||||
Authors | Chen, M. / Pintilie, G. | ||||||
Funding support | United States, 1items
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Citation | Journal: Nat Methods / Year: 2024 Title: Improving resolution and resolvability of single-particle cryoEM structures using Gaussian mixture models. Authors: Muyuan Chen / Michael F Schmid / Wah Chiu / Abstract: Cryogenic electron microscopy is widely used in structural biology, but its resolution is often limited by the dynamics of the macromolecule. Here we developed a refinement protocol based on Gaussian ...Cryogenic electron microscopy is widely used in structural biology, but its resolution is often limited by the dynamics of the macromolecule. Here we developed a refinement protocol based on Gaussian mixture models that integrates particle orientation and conformation estimation and improves the alignment for flexible domains of protein structures. We demonstrated this protocol on multiple datasets, resulting in improved resolution and resolvability, locally and globally, by visual and quantitative measures. | ||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8u2c.cif.gz | 352 KB | Display | PDBx/mmCIF format |
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PDB format | pdb8u2c.ent.gz | 277.9 KB | Display | PDB format |
PDBx/mmJSON format | 8u2c.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/u2/8u2c ftp://data.pdbj.org/pub/pdb/validation_reports/u2/8u2c | HTTPS FTP |
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-Related structure data
Related structure data | 41845MC 8u26C 8u28C M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
Experimental dataset #1 | Data reference: 10.6019/EMPIAR-10374 / Data set type: EMPIAR |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 72385.852 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: ABCG2, ABCP, BCRP, BCRP1, MXR / Production host: Homo sapiens (human) References: UniProt: Q9UNQ0, ABC-type xenobiotic transporter #2: Antibody | Mass: 23594.016 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Production host: Mus musculus (house mouse) #3: Antibody | Mass: 23843.633 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Mus musculus (house mouse) / Production host: Mus musculus (house mouse) #4: Antibody | Mass: 22986.840 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Details: The Fab domains were not resolved in the original structure and, therefore, were not modeled. With the implementation of an improved method, we can now resolve the domains within the CryoEM ...Details: The Fab domains were not resolved in the original structure and, therefore, were not modeled. With the implementation of an improved method, we can now resolve the domains within the CryoEM density map. However, given the absence of FAB information in the original structure, we opted to fit a representative FAB structure, utilizing PDB ID 7FAB as a reference. Source: (gene. exp.) Homo sapiens (human) / Production host: Homo sapiens (human) #5: Antibody | Mass: 22866.355 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Details: The Fab domains were not resolved in the original structure and, therefore, were not modeled. With the implementation of an improved method, we can now resolve the domains within the CryoEM ...Details: The Fab domains were not resolved in the original structure and, therefore, were not modeled. With the implementation of an improved method, we can now resolve the domains within the CryoEM density map. However, given the absence of FAB information in the original structure, we opted to fit a representative FAB structure, utilizing PDB ID 7FAB as a reference. Source: (gene. exp.) Homo sapiens (human) / Production host: Homo sapiens (human) |
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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: inhibitor-bound ABCG2 / Type: COMPLEX / Details: Re-refinement from EMPIAR-10374 / Entity ID: all / Source: RECOMBINANT |
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Source (natural) | Organism: Homo sapiens (human) |
Source (recombinant) | Organism: Homo sapiens (human) |
Buffer solution | pH: 7.5 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 2000 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 30 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING ONLY | |||||||||||||||||||||
Symmetry | Point symmetry: C2 (2 fold cyclic) | |||||||||||||||||||||
3D reconstruction | Resolution: 2.5 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 284831 / Symmetry type: POINT | |||||||||||||||||||||
Atomic model building |
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