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- PDB-4am7: ADP-BOUND C-TERMINAL DOMAIN OF ACTIN-RELATED PROTEIN ARP8 FROM S.... -

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Basic information

Entry
Database: PDB / ID: 4am7
TitleADP-BOUND C-TERMINAL DOMAIN OF ACTIN-RELATED PROTEIN ARP8 FROM S. CEREVISIAE
ComponentsACTIN-LIKE PROTEIN ARP8
KeywordsNUCLEAR PROTEIN / CHROMATIN REMODELLING COMPLEX / ATP-BINDING PROTEIN / NUCLEAR ACTIN-RELATED PROTEIN / TRANSCRIPTION REGULATION / DNA REPAIR
Function / homology
Function and homology information


Ino80 complex / mitotic recombination / double-strand break repair / cytoskeleton / chromatin remodeling / DNA repair / mRNA binding / DNA damage response / regulation of DNA-templated transcription / ATP binding ...Ino80 complex / mitotic recombination / double-strand break repair / cytoskeleton / chromatin remodeling / DNA repair / mRNA binding / DNA damage response / regulation of DNA-templated transcription / ATP binding / nucleus / cytoplasm
Similarity search - Function
Nucleotidyltransferase; domain 5 - #580 / ATPase, substrate binding domain, subdomain 4 / Actin; Chain A, domain 4 / ATPase, nucleotide binding domain / Actin / Actin family / Actin / ATPase, nucleotide binding domain / Nucleotidyltransferase; domain 5 / Alpha-Beta Complex ...Nucleotidyltransferase; domain 5 - #580 / ATPase, substrate binding domain, subdomain 4 / Actin; Chain A, domain 4 / ATPase, nucleotide binding domain / Actin / Actin family / Actin / ATPase, nucleotide binding domain / Nucleotidyltransferase; domain 5 / Alpha-Beta Complex / 2-Layer Sandwich / Alpha Beta
Similarity search - Domain/homology
ADENOSINE-5'-DIPHOSPHATE / Actin-like protein ARP8
Similarity search - Component
Biological speciesSACCHAROMYCES CEREVISIAE (brewer's yeast)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.25 Å
AuthorsWuerges, J. / Saravanan, M. / Bose, D. / Cook, N.J. / Zhang, X. / Wigley, D.B.
CitationJournal: Proc Natl Acad Sci U S A / Year: 2012
Title: Interactions between the nucleosome histone core and Arp8 in the INO80 chromatin remodeling complex.
Authors: Matheshwaran Saravanan / Jochen Wuerges / Daniel Bose / Elizabeth A McCormack / Nicola J Cook / Xiaodong Zhang / Dale B Wigley /
Abstract: Actin-related protein Arp8 is a component of the INO80 chromatin remodeling complex. Yeast Arp8 (yArp8) comprises two domains: a 25-KDa N-terminal domain, found only in yeast, and a 75-KDa C-terminal ...Actin-related protein Arp8 is a component of the INO80 chromatin remodeling complex. Yeast Arp8 (yArp8) comprises two domains: a 25-KDa N-terminal domain, found only in yeast, and a 75-KDa C-terminal domain (yArp8CTD) that contains the actin fold and is conserved across other species. The crystal structure shows that yArp8CTD contains three insertions within the actin core. Using a combination of biochemistry and EM, we show that Arp8 forms a complex with nucleosomes, and that the principal interactions are via the H3 and H4 histones, mediated through one of the yArp8 insertions. We show that recombinant yArp8 exists in monomeric and dimeric states, but the dimer is the biologically relevant form required for stable interactions with histones that exploits the twofold symmetry of the nucleosome core. Taken together, these data provide unique insight into the stoichiometry, architecture, and molecular interactions between components of the INO80 remodeling complex and nucleosomes, providing a first step toward building up the structure of the complex.
History
DepositionMar 7, 2012Deposition site: PDBE / Processing site: PDBE
Revision 1.0Dec 12, 2012Provider: repository / Type: Initial release
Revision 1.1Jan 16, 2013Group: Database references
Revision 1.2Nov 13, 2019Group: Data collection / Database references / Other / Category: pdbx_database_related / pdbx_database_status
Item: _pdbx_database_related.content_type / _pdbx_database_status.status_code_sf
Revision 1.3Dec 20, 2023Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model / struct_ncs_dom_lim / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _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 / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: ACTIN-LIKE PROTEIN ARP8
B: ACTIN-LIKE PROTEIN ARP8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)150,4814
Polymers149,6272
Non-polymers8542
Water00
1
A: ACTIN-LIKE PROTEIN ARP8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)75,2412
Polymers74,8131
Non-polymers4271
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: ACTIN-LIKE PROTEIN ARP8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)75,2412
Polymers74,8131
Non-polymers4271
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)137.718, 89.795, 149.647
Angle α, β, γ (deg.)90.00, 114.37, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B

NCS domain segments:

Ens-ID: 1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ASNASNTRPTRP5AA263 - 30137 - 75
21ASNASNTRPTRP5BB263 - 30137 - 75
12LEULEUGLUGLU6AA302 - 31876 - 92
22LEULEUGLUGLU6BB302 - 31876 - 92
13GLNGLNMETMET5AA319 - 33893 - 112
23GLNGLNMETMET5BB319 - 33893 - 112
14ARGARGARGARG6AA339 - 391113 - 165
24ARGARGARGARG6BB339 - 391113 - 165
15CYSCYSPROPRO5AA392 - 442166 - 216
25CYSCYSPROPRO5BB392 - 442166 - 216
16THRTHRCYSCYS4AA443 - 489217 - 263
26THRTHRCYSCYS4BB443 - 489217 - 263
17TYRTYRTHRTHR6AA490 - 498264 - 272
27TYRTYRTHRTHR6BB490 - 498264 - 272
18CYSCYSSERSER4AA499 - 555273 - 329
28CYSCYSSERSER4BB499 - 555273 - 329
19LYSLYSPHEPHE5AA556 - 584330 - 358
29LYSLYSPHEPHE5BB556 - 584330 - 358
110METMETGLUGLU6AA585 - 594359 - 368
210METMETGLUGLU6BB585 - 594359 - 368
111LYSLYSGLNGLN5AA595 - 686369 - 460
211LYSLYSGLNGLN5BB595 - 686369 - 460
112ASPASPLEULEU6AA687 - 696461 - 470
212ASPASPLEULEU6BB687 - 696461 - 470
113LYSLYSSERSER5AA697 - 759471 - 533
213LYSLYSSERSER5BB697 - 759471 - 533
114SERSERHISHIS6AA760 - 823534 - 597
214SERSERHISHIS6BB760 - 823534 - 597
115ILEILEASNASN6AA824 - 837598 - 611
215ILEILEASNASN6BB824 - 837598 - 611
116PROPROTYRTYR5AA838 - 881612 - 655
216PROPROTYRTYR5BB838 - 881612 - 655

NCS oper: (Code: given
Matrix: (0.67099, 0.08449, 0.73663), (0.72002, -0.31144, -0.62014), (0.17702, 0.9465, -0.26981)
Vector: -46.07872, 105.46299, -34.37724)

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Components

#1: Protein ACTIN-LIKE PROTEIN ARP8


Mass: 74813.469 Da / Num. of mol.: 2 / Fragment: C-TERMINAL DOMAIN, RESIDUES 248-881
Source method: isolated from a genetically manipulated source
Details: ADP-BOUND STATE OF THE PROTEIN
Source: (gene. exp.) SACCHAROMYCES CEREVISIAE (brewer's yeast)
Strain: W303-1A / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): B834 / Variant (production host): PLYSS / References: UniProt: Q12386
#2: Chemical ChemComp-ADP / ADENOSINE-5'-DIPHOSPHATE


Mass: 427.201 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C10H15N5O10P2 / Comment: ADP, energy-carrying molecule*YM

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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: 2.8 Å3/Da / Density % sol: 57 % / Description: NONE
Crystal growpH: 6.5
Details: 19.5% W/V PEG 5000 MME, 0.32 M LI2SO4, 0.1 M MES PH 6.5, 0.002 M ADP, 0.005 M MGCL2

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9763
DetectorType: ADSC CCD / Detector: CCD / Date: Dec 2, 2009 / Details: MIRRORS
RadiationMonochromator: DOUBLE CRYSTAL / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 3.25→20 Å / Num. obs: 25357 / % possible obs: 96.4 % / Observed criterion σ(I): 2 / Redundancy: 3.1 % / Biso Wilson estimate: 96.105 Å2 / Rmerge(I) obs: 0.09 / Net I/σ(I): 8.5
Reflection shellResolution: 3.25→3.43 Å / Redundancy: 2.8 % / Rmerge(I) obs: 0.44 / Mean I/σ(I) obs: 2.5 / % possible all: 94.6

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Processing

Software
NameVersionClassification
REFMAC5.5.0088refinement
MOSFLMdata reduction
SCALAdata scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 4AM6
Resolution: 3.25→19.81 Å / Cor.coef. Fo:Fc: 0.932 / Cor.coef. Fo:Fc free: 0.874 / SU B: 63.316 / SU ML: 0.506 / Cross valid method: THROUGHOUT / ESU R: 1.863 / ESU R Free: 0.617 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. N-TERMINAL TAG AND RESIDUES 248 TO 258 OF ARP8 HAVE NOT BEEN LOCATED IN THE ELECTRON DENSITY MAP.
RfactorNum. reflection% reflectionSelection details
Rfree0.28417 1301 5.1 %RANDOM
Rwork0.2192 ---
obs0.22259 24054 96.45 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 121.118 Å2
Baniso -1Baniso -2Baniso -3
1--1.61 Å20 Å20.62 Å2
2--1.81 Å20 Å2
3---0.31 Å2
Refinement stepCycle: LAST / Resolution: 3.25→19.81 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms10072 0 54 0 10126
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.02210356
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.5661.96314058
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg7.28151244
X-RAY DIFFRACTIONr_dihedral_angle_2_deg41.10225.412510
X-RAY DIFFRACTIONr_dihedral_angle_3_deg23.055151838
X-RAY DIFFRACTIONr_dihedral_angle_4_deg22.0251542
X-RAY DIFFRACTIONr_chiral_restr0.1060.21566
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0217840
X-RAY DIFFRACTIONr_gen_planes_other
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_it0.5771.56250
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it1.088210196
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it1.22234106
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it2.1594.53862
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Ens-ID: 1 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1A2173medium positional0.370.5
2B2173medium positional0.370.5
1A2830loose positional1.345
2B2830loose positional1.345
1A2173medium thermal4.172
2B2173medium thermal4.172
1A2830loose thermal3.4510
2B2830loose thermal3.4510
LS refinement shellResolution: 3.25→3.332 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.333 87 -
Rwork0.284 1611 -
obs--89.89 %
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
14.5968-1.2268-0.03513.97480.28973.910.1739-0.07530.1497-0.0428-0.41430.3213-0.002-0.23610.24040.0110.0009-0.00820.0758-0.10350.2317-115.947.9272-9.2815
23.6602-0.5129-0.47765.5995-0.6174.61870.13870.57-0.2882-0.47080.12890.30620.3093-0.09-0.26760.2250.1903-0.06090.3292-0.01820.0874-129.958624.9787-44.972
30.33420.93911.09155.3386-0.06869.04530.04380.06810.027-0.377-0.33690.0270.30420.26150.29310.20880.2421-0.03810.2983-0.0570.2405-113.3123.7301-33.9293
49.5123-0.8758-1.33542.9228-1.56241.39520.08340.49870.25340.03830.14930.413-0.1795-0.1966-0.23280.29410.21330.06780.31980.22280.2268-145.97244.4025-40.9163
56.3502-2.883-0.00323.0802-0.05463.816-0.0703-0.3585-0.23070.3221-0.24460.13650.18850.22240.3150.0662-0.06440.00140.16460.10630.2131-93.7639-4.4049-1.5198
63.83930.8693-0.53097.6439-0.85173.0049-0.16530.89620.0349-0.82440.1769-0.77550.01480.2496-0.01160.39150.19570.19830.65740.05930.1347-110.484339.9475-55.1302
73.88720.90311.00836.38980.5834.27730.16640.7933-0.5715-0.5669-0.27640.08030.10120.10910.10990.21940.18960.07890.2811-0.05390.2664-97.1448-14.6265-26.8138
85.43970.33052.01012.2378-0.91863.5001-0.11161.00341.1387-0.45270.1960.3524-0.3511-0.0631-0.08440.62010.24240.09430.70010.49830.4347-132.349856.9903-58.518
95.30030.17243.46352.2014-1.01225.6650.20930.413-0.6462-0.7217-0.357-0.20740.26220.07810.14770.44480.24960.11350.1984-0.07010.3446-95.5903-14.9124-28.2492
109.03662.7632-1.17132.337-0.48212.1029-0.08420.54850.0015-0.51820.06430.1885-0.65730.09870.01990.76770.29990.10650.5730.37110.6418-132.437658.9605-57.9273
111.8428-1.8215-0.10943.60670.58371.35840.22080.31410.6147-0.3866-0.5445-0.6085-0.66020.53840.32370.3766-0.2437-0.03710.42020.22550.5855-89.228913.537-8.1554
124.79440.1076-2.03821.9002-0.05781.1160.2527-0.78280.62260.7544-0.0073-0.5095-0.29020.6166-0.24540.70530.0357-0.07690.7416-0.00970.3672-110.852841.8178-35.3126
139.35411.714-1.61318.9833-0.99696.9434-0.1572-1.0950.62611.12790.2804-0.545-0.14781.0419-0.12320.2704-0.1179-0.28420.76320.07630.4864-74.22778.530416.4896
141.25943.07320.73247.72311.81260.43350.30070.2463-0.38950.8616-0.0914-0.94270.24350.0992-0.20930.68560.21910.14022.10270.36441.3038-83.281639.6853-42.7209
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A259 - 293
2X-RAY DIFFRACTION1A403 - 494
3X-RAY DIFFRACTION1A836 - 881
4X-RAY DIFFRACTION2B259 - 293
5X-RAY DIFFRACTION2B403 - 494
6X-RAY DIFFRACTION2B836 - 881
7X-RAY DIFFRACTION3A294 - 305
8X-RAY DIFFRACTION3A381 - 402
9X-RAY DIFFRACTION4B294 - 305
10X-RAY DIFFRACTION4B381 - 402
11X-RAY DIFFRACTION5A495 - 529
12X-RAY DIFFRACTION5A700 - 759
13X-RAY DIFFRACTION5A824 - 835
14X-RAY DIFFRACTION6B495 - 529
15X-RAY DIFFRACTION6B700 - 759
16X-RAY DIFFRACTION6B824 - 835
17X-RAY DIFFRACTION7A530 - 619
18X-RAY DIFFRACTION8B530 - 619
19X-RAY DIFFRACTION9A306 - 380
20X-RAY DIFFRACTION10B306 - 380
21X-RAY DIFFRACTION11A620 - 699
22X-RAY DIFFRACTION12B620 - 699
23X-RAY DIFFRACTION13A760 - 823
24X-RAY DIFFRACTION14B760 - 823

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