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- PDB-5tqq: Cryo-electron microscopy structure of a bovine CLC-K chloride cha... -

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Entry
Database: PDB / ID: 5tqq
TitleCryo-electron microscopy structure of a bovine CLC-K chloride channel, main (class 1) conformation
DescriptorChloride channel protein
Fab fragment
light chain
Fab fragment
heavy chain
(Monoclonal ...) x 2
KeywordsTRANSPORT PROTEIN / CLC / chloride channel / membrane / kidney
Specimen sourceBos taurus / mammal / Bovine / ウシ /
Mus musculus / mammal / house mouse / ハツカネズミ, はつかねずみ /
MethodElectron microscopy (3.76 A resolution / Single particle)
AuthorsPark, E. / MacKinnon, R.
CitationNature, 2017, 541, 500-505

Nature, 2017, 541, 500-505 StrPapers
Structure of a CLC chloride ion channel by cryo-electron microscopy.
Eunyong Park / Ernest B Campbell / Roderick MacKinnon

DateDeposition: Oct 24, 2016 / Release: Jan 4, 2017 / Last modification: Feb 8, 2017

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Assembly

Deposited unit
A: Chloride channel protein
L: Monoclonal antibody, Fab fragment, light chain
H: Monoclonal antibody, Fab fragment, heavy chain
B: Chloride channel protein
M: Monoclonal antibody, Fab fragment, light chain
I: Monoclonal antibody, Fab fragment, heavy chain


Theoretical massNumber of molelcules
Total (without water)195,9576
Polyers195,9576
Non-polymers00
Water0
#1


TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA

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Components

#1: Polypeptide(L)Chloride channel protein


Mass: 73521.484 Da / Num. of mol.: 2 / Mutation: N373Q / Source: (gene. exp.) Bos taurus / mammal / ウシ / / References: UniProt: E1B792

Cellular component

Molecular function

Biological process

#2: Polypeptide(L)Monoclonal antibody, Fab fragment, light chain


Mass: 11824.102 Da / Num. of mol.: 2
Source: (gene. exp.) Mus musculus / mammal / ハツカネズミ, はつかねずみ /
#3: Polypeptide(L)Monoclonal antibody, Fab fragment, heavy chain


Mass: 12633.068 Da / Num. of mol.: 2
Source: (gene. exp.) Mus musculus / mammal / ハツカネズミ, はつかねずみ /

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentReconstruction method: SINGLE PARTICLE

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Sample preparation

ComponentName: A bovine CLC-K channel complexed with a monoclonal antibody fragment (Fab)
Details: Fab fragment generated by proteolytic (papain) cleavage of murine IgG antibody.
Type: COMPLEX
Specimen supportGrid material: COPPER / Grid type: Quantifoil R 1.2/1.3
VitrificationInstrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 90 %

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyMicroscope model: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD

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Processing

SoftwareName: REFMAC / Version: 5.8.0088 / Classification: refinement
ComputingStructure refinement: REFMAC 5.8.0088
3D reconstructionResolution: 3.76 / Resolution method: FSC 0.143 CUT-OFF / Number of particles: 82167
Details: the indicated resolution was estimated on the primary map using the provided mask including Fab.

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