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- PDB-6qqp: Aplysia californica AChBP in complex with 2-Fluoro-(carbamoylpyri... -

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Basic information

Entry
Database: PDB / ID: 6qqp
TitleAplysia californica AChBP in complex with 2-Fluoro-(carbamoylpyridinyl)deschloroepibatidine analogue (2)
ComponentsSoluble acetylcholine receptor
KeywordsSIGNALING PROTEIN / AChBP / Acetylcholine
Function / homology
Function and homology information


extracellular ligand-gated monoatomic ion channel activity / transmembrane signaling receptor activity / membrane / identical protein binding / metal ion binding
Similarity search - Function
Acetylcholine Binding Protein; Chain: A, / Neurotransmitter-gated ion-channel ligand-binding domain / Neurotransmitter-gated ion-channel / Neurotransmitter-gated ion-channel ligand-binding domain / Neurotransmitter-gated ion-channel ligand-binding domain superfamily / Neurotransmitter-gated ion-channel ligand binding domain / Distorted Sandwich / Mainly Beta
Similarity search - Domain/homology
Chem-JCW / Soluble acetylcholine receptor
Similarity search - Component
Biological speciesAplysia californica (California sea hare)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.4 Å
AuthorsBueno, R.V. / Davis, S. / Dawson, A. / Hunter, W.N.
CitationJournal: Acta Crystallogr.,Sect.D / Year: 2022
Title: Interactions between 2'-fluoro-(carbamoylpyridinyl)deschloroepibatidine analogues and acetylcholine-binding protein inform on potent antagonist activity against nicotinic receptors
Authors: Bueno, R.V. / Davis, S. / Dawson, A. / Ondachi, P.W. / Carroll, F.I. / Hunter, W.N.
History
DepositionFeb 18, 2019Deposition site: PDBE / Processing site: PDBE
Revision 1.0Mar 18, 2020Provider: repository / Type: Initial release
Revision 1.1Jul 29, 2020Group: Data collection / Derived calculations / Structure summary
Category: chem_comp / entity ...chem_comp / entity / pdbx_chem_comp_identifier / pdbx_entity_nonpoly / struct_conn / struct_site / struct_site_gen
Item: _chem_comp.name / _entity.pdbx_description ..._chem_comp.name / _entity.pdbx_description / _pdbx_entity_nonpoly.name / _struct_conn.pdbx_role
Description: Carbohydrate remediation / Provider: repository / Type: Remediation
Revision 1.2Mar 2, 2022Group: Database references / Structure summary
Category: chem_comp / citation ...chem_comp / citation / citation_author / database_2
Item: _chem_comp.pdbx_synonyms / _citation.country ..._chem_comp.pdbx_synonyms / _citation.country / _citation.journal_abbrev / _citation.journal_id_ASTM / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.pdbx_database_id_DOI / _citation.title / _citation.year / _database_2.pdbx_DOI / _database_2.pdbx_database_accession
Revision 1.3Jan 24, 2024Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model / struct_ncs_dom_lim
Item: _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id ..._struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Soluble acetylcholine receptor
B: Soluble acetylcholine receptor
C: Soluble acetylcholine receptor
D: Soluble acetylcholine receptor
E: Soluble acetylcholine receptor
F: Soluble acetylcholine receptor
G: Soluble acetylcholine receptor
H: Soluble acetylcholine receptor
I: Soluble acetylcholine receptor
J: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)292,32790
Polymers283,26810
Non-polymers9,06080
Water16,646924
1
A: Soluble acetylcholine receptor
B: Soluble acetylcholine receptor
C: Soluble acetylcholine receptor
D: Soluble acetylcholine receptor
E: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)146,22646
Polymers141,6345
Non-polymers4,59241
Water905
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
F: Soluble acetylcholine receptor
G: Soluble acetylcholine receptor
H: Soluble acetylcholine receptor
I: Soluble acetylcholine receptor
J: Soluble acetylcholine receptor
hetero molecules


Theoretical massNumber of molelcules
Total (without water)146,10144
Polymers141,6345
Non-polymers4,46839
Water905
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)209.530, 136.948, 131.552
Angle α, β, γ (deg.)90.00, 102.74, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23D
14A
24E
15A
25F
16A
26G
17A
27H
18A
28I
19A
29J
110B
210C
111B
211D
112B
212E
113B
213F
114B
214G
115B
215H
116B
216I
117B
217J
118C
218D
119C
219E
120C
220F
121C
221G
122C
222H
123C
223I
124C
224J
125D
225E
126D
226F
127D
227G
128D
228H
129D
229I
130D
230J
131E
231F
132E
232G
133E
233H
134E
234I
135E
235J
136F
236G
137F
237H
138F
238I
139F
239J
140G
240H
141G
241I
142G
242J
143H
243I
144H
244J
145I
245J

NCS domain segments:

Component-ID: 0 / Beg auth comp-ID: GLN / Beg label comp-ID: GLN / Refine code: 0

Dom-IDEns-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ARGARGAA20 - 22420 - 224
21ARGARGBB20 - 22420 - 224
12ARGARGAA20 - 22420 - 224
22ARGARGCC20 - 22420 - 224
13ARGARGAA20 - 22420 - 224
23ARGARGDD20 - 22420 - 224
14GLUGLUAA20 - 22320 - 223
24GLUGLUEE20 - 22320 - 223
15ARGARGAA20 - 22420 - 224
25ARGARGFF20 - 22420 - 224
16ARGARGAA20 - 22420 - 224
26ARGARGGG20 - 22420 - 224
17ARGARGAA20 - 22420 - 224
27ARGARGHH20 - 22420 - 224
18ARGARGAA20 - 22420 - 224
28ARGARGII20 - 22420 - 224
19ARGARGAA20 - 22420 - 224
29ARGARGJJ20 - 22420 - 224
110ARGARGBB20 - 22420 - 224
210ARGARGCC20 - 22420 - 224
111ARGARGBB20 - 22420 - 224
211ARGARGDD20 - 22420 - 224
112GLUGLUBB20 - 22320 - 223
212GLUGLUEE20 - 22320 - 223
113ARGARGBB20 - 22420 - 224
213ARGARGFF20 - 22420 - 224
114ARGARGBB20 - 22420 - 224
214ARGARGGG20 - 22420 - 224
115ARGARGBB20 - 22420 - 224
215ARGARGHH20 - 22420 - 224
116ARGARGBB20 - 22420 - 224
216ARGARGII20 - 22420 - 224
117ARGARGBB20 - 22420 - 224
217ARGARGJJ20 - 22420 - 224
118ARGARGCC20 - 22420 - 224
218ARGARGDD20 - 22420 - 224
119GLUGLUCC20 - 22320 - 223
219GLUGLUEE20 - 22320 - 223
120ARGARGCC20 - 22420 - 224
220ARGARGFF20 - 22420 - 224
121ARGARGCC20 - 22420 - 224
221ARGARGGG20 - 22420 - 224
122ARGARGCC20 - 22420 - 224
222ARGARGHH20 - 22420 - 224
123ARGARGCC20 - 22420 - 224
223ARGARGII20 - 22420 - 224
124ARGARGCC20 - 22420 - 224
224ARGARGJJ20 - 22420 - 224
125GLUGLUDD20 - 22320 - 223
225GLUGLUEE20 - 22320 - 223
126ARGARGDD20 - 22420 - 224
226ARGARGFF20 - 22420 - 224
127ARGARGDD20 - 22420 - 224
227ARGARGGG20 - 22420 - 224
128ARGARGDD20 - 22420 - 224
228ARGARGHH20 - 22420 - 224
129ARGARGDD20 - 22420 - 224
229ARGARGII20 - 22420 - 224
130ARGARGDD20 - 22420 - 224
230ARGARGJJ20 - 22420 - 224
131GLUGLUEE20 - 22320 - 223
231GLUGLUFF20 - 22320 - 223
132GLUGLUEE20 - 22320 - 223
232GLUGLUGG20 - 22320 - 223
133GLUGLUEE20 - 22320 - 223
233GLUGLUHH20 - 22320 - 223
134GLUGLUEE20 - 22320 - 223
234GLUGLUII20 - 22320 - 223
135GLUGLUEE20 - 22320 - 223
235GLUGLUJJ20 - 22320 - 223
136ARGARGFF20 - 22420 - 224
236ARGARGGG20 - 22420 - 224
137ARGARGFF20 - 22420 - 224
237ARGARGHH20 - 22420 - 224
138ARGARGFF20 - 22420 - 224
238ARGARGII20 - 22420 - 224
139ARGARGFF20 - 22420 - 224
239ARGARGJJ20 - 22420 - 224
140ARGARGGG20 - 22420 - 224
240ARGARGHH20 - 22420 - 224
141ARGARGGG20 - 22420 - 224
241ARGARGII20 - 22420 - 224
142ARGARGGG20 - 22420 - 224
242ARGARGJJ20 - 22420 - 224
143ARGARGHH20 - 22420 - 224
243ARGARGII20 - 22420 - 224
144ARGARGHH20 - 22420 - 224
244ARGARGJJ20 - 22420 - 224
145ARGARGII20 - 22420 - 224
245ARGARGJJ20 - 22420 - 224

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45

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Components

#1: Protein
Soluble acetylcholine receptor


Mass: 28326.750 Da / Num. of mol.: 10
Source method: isolated from a genetically manipulated source
Details: The sequence conflicts are because we have used the AChBP gene sequence provided in the Aplysia californica genome (http://genome-euro.ucsc.edu/cgi-bin/hgGateway?hgsid=589127391_ ...Details: The sequence conflicts are because we have used the AChBP gene sequence provided in the Aplysia californica genome (http://genome-euro.ucsc.edu/cgi-bin/hgGateway?hgsid=589127391_t0Vv5jEv8KyWGt7MjPEaZKNxTL3u&redirect=manual&source=genome.cse.ucsc.edu), which differs from the UniProt Q8WSF8 Sequence by identifying both ALA 60 and and ALA 155 as VAL.
Source: (gene. exp.) Aplysia californica (California sea hare)
Cell line (production host): Sf9 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: Q8WSF8
#2: Chemical
ChemComp-JCW / 2-[5-[(1~{R},2~{R},4~{S})-7-azabicyclo[2.2.1]heptan-2-yl]-2-fluoranyl-pyridin-3-yl]pyridine-4-carboxamide


Mass: 312.341 Da / Num. of mol.: 10 / Source method: obtained synthetically / Formula: C17H17FN4O / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical...
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 60 / Source method: obtained synthetically / Formula: C2H6O2
#4: Sugar
ChemComp-NAG / 2-acetamido-2-deoxy-beta-D-glucopyranose / N-acetyl-beta-D-glucosamine / 2-acetamido-2-deoxy-beta-D-glucose / 2-acetamido-2-deoxy-D-glucose / 2-acetamido-2-deoxy-glucose / N-ACETYL-D-GLUCOSAMINE / N-Acetylglucosamine


Type: D-saccharide, beta linking / Mass: 221.208 Da / Num. of mol.: 10
Source method: isolated from a genetically manipulated source
Formula: C8H15NO6
IdentifierTypeProgram
DGlcpNAcbCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
N-acetyl-b-D-glucopyranosamineCOMMON NAMEGMML 1.0
b-D-GlcpNAcIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcNAcSNFG CARBOHYDRATE SYMBOLGMML 1.0
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 924 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 3.49 Å3/Da / Density % sol: 64.77 % / Description: Block
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 4.2
Details: Reservoir: 0.2 M NaCl, 0.1 M Phosphate/citrate pH 4.2 8% PEG 8000 Buffer: 50 mM Tris 250 mM NaCl pH 7.5 Protein concentration: 4 mg/ml Microseeded

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I24 / Wavelength: 0.96858 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Jul 3, 2017
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.96858 Å / Relative weight: 1
ReflectionResolution: 2.4→46.82 Å / Num. obs: 139199 / % possible obs: 98.7 % / Redundancy: 2.9 % / Biso Wilson estimate: 29 Å2 / CC1/2: 0.95 / Rmerge(I) obs: 0.089 / Net I/σ(I): 8.2
Reflection shellResolution: 2.4→2.44 Å / Redundancy: 2.8 % / Rmerge(I) obs: 0.51 / Mean I/σ(I) obs: 2.5 / Num. unique obs: 6884 / CC1/2: 0.65 / % possible all: 98.7

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Processing

Software
NameVersionClassification
REFMAC5.8.0189refinement
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 5o87
Resolution: 2.4→46.82 Å / Cor.coef. Fo:Fc: 0.93 / Cor.coef. Fo:Fc free: 0.915 / SU B: 6.257 / SU ML: 0.138 / Cross valid method: THROUGHOUT / ESU R: 0.253 / ESU R Free: 0.191 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.21003 6903 5 %RANDOM
Rwork0.18785 ---
obs0.18893 131926 98.25 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 34.364 Å2
Baniso -1Baniso -2Baniso -3
1--2.47 Å20 Å2-0.22 Å2
2--3.82 Å20 Å2
3----1.14 Å2
Refinement stepCycle: LAST / Resolution: 2.4→46.82 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms16388 0 610 924 17922
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0110.0217520
X-RAY DIFFRACTIONr_bond_other_d0.0010.0215555
X-RAY DIFFRACTIONr_angle_refined_deg1.491.96323864
X-RAY DIFFRACTIONr_angle_other_deg0.9193.00136239
X-RAY DIFFRACTIONr_dihedral_angle_1_deg8.0645.0142089
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.61224.62803
X-RAY DIFFRACTIONr_dihedral_angle_3_deg13.479152753
X-RAY DIFFRACTIONr_dihedral_angle_4_deg16.1941591
X-RAY DIFFRACTIONr_chiral_restr0.0840.22664
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.02119865
X-RAY DIFFRACTIONr_gen_planes_other0.0010.023490
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it2.2643.258251
X-RAY DIFFRACTIONr_mcbond_other2.2643.258252
X-RAY DIFFRACTIONr_mcangle_it3.6384.86910318
X-RAY DIFFRACTIONr_mcangle_other3.6384.86910318
X-RAY DIFFRACTIONr_scbond_it3.3073.7359268
X-RAY DIFFRACTIONr_scbond_other3.3073.7369269
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other5.3885.44713526
X-RAY DIFFRACTIONr_long_range_B_refined7.43738.06918328
X-RAY DIFFRACTIONr_long_range_B_other7.39737.99418226
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A130980.03
12B130980.03
21A131320.04
22C131320.04
31A131220.03
32D131220.03
41A130100.04
42E130100.04
51A131800.03
52F131800.03
61A131480.04
62G131480.04
71A131020.04
72H131020.04
81A131600.03
82I131600.03
91A131460.03
92J131460.03
101B131620.04
102C131620.04
111B131340.04
112D131340.04
121B130080.04
122E130080.04
131B131640.03
132F131640.03
141B131260.05
142G131260.05
151B130540.04
152H130540.04
161B131480.04
162I131480.04
171B131440.03
172J131440.03
181C131040.04
182D131040.04
191C129860.04
192E129860.04
201C131360.04
202F131360.04
211C131160.05
212G131160.05
221C130340.05
222H130340.05
231C131280.04
232I131280.04
241C131100.04
242J131100.04
251D129980.04
252E129980.04
261D131840.03
262F131840.03
271D131480.04
272G131480.04
281D130880.04
282H130880.04
291D131960.03
292I131960.03
301D131840.02
302J131840.02
311E129900.04
312F129900.04
321E129760.04
322G129760.04
331E129380.04
332H129380.04
341E129920.04
342I129920.04
351E129880.04
352J129880.04
361F131560.04
362G131560.04
371F131100.04
372H131100.04
381F131460.04
382I131460.04
391F131920.02
392J131920.02
401G130560.04
402H130560.04
411G131600.04
412I131600.04
421G131080.04
422J131080.04
431H131580.04
432I131580.04
441H131440.04
442J131440.04
451I131240.04
452J131240.04
LS refinement shellResolution: 2.4→2.462 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.309 519 -
Rwork0.286 9737 -
obs--98.65 %

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