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- PDB-2x7l: Implications of the HIV-1 Rev dimer structure at 3.2A resolution ... -

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Basic information

Entry
Database: PDB / ID: 2x7l
TitleImplications of the HIV-1 Rev dimer structure at 3.2A resolution for multimeric binding to the Rev response element
Components
  • FAB HEAVY CHAIN
  • FAB LIGHT CHAIN
  • PROTEIN REV
KeywordsIMMUNE SYSTEM / NUCLEAR EXPORT / POST-TRANSCRIPTIONAL REGULATION
Function / homology
Function and homology information


host cell nucleolus / mRNA transport / viral process / protein export from nucleus / host cell cytoplasm / DNA-binding transcription factor activity / RNA binding
Similarity search - Function
Helix Hairpins - #630 / Anti-repression trans-activator protein, REV protein / REV protein (anti-repression trans-activator protein) / Helix Hairpins / Helix non-globular / Special / Immunoglobulins / Immunoglobulin-like / Sandwich / Mainly Beta
Similarity search - Domain/homology
Biological speciessynthetic construct (others)
HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 BH10
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.17 Å
AuthorsDiMattia, M.A. / Watts, N.R. / Stahl, S.J. / Rader, C. / Wingfield, P.T. / Stuart, D.I. / Steven, A.C. / Grimes, J.M.
Citation
Journal: Proc.Natl.Acad.Sci.USA / Year: 2010
Title: Implications of the HIV-1 Rev Dimer Structure at 3. 2 A Resolution for Multimeric Binding to the Rev Response Element.
Authors: Dimattia, M.A. / Watts, N.R. / Stahl, S.J. / Rader, C. / Wingfield, P.T. / Stuart, D.I. / Steven, A.C. / Grimes, J.M.
#1: Journal: J.Mol.Biol. / Year: 2010
Title: Generation and Characterization of a Chimeric Rabbit/Human Fab for Co-Crystallization of HIV-1 Rev.
Authors: Stahl, S.J. / Watts, N.R. / Rader, C. / Dimattia, M.A. / Mage, R.G. / Palmer, I. / Kaufman, J.D. / Grimes, J.M. / Stuart, D.I. / Steven, A.C. / Wingfield, P.T.
History
DepositionMar 1, 2010Deposition site: PDBE / Processing site: PDBE
Revision 1.0Mar 23, 2010Provider: repository / Type: Initial release
Revision 1.1Sep 18, 2013Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description / Source and taxonomy / Structure summary / Version format compliance
Revision 1.2Feb 20, 2019Group: Data collection / Other / Source and taxonomy
Category: entity_src_gen / pdbx_database_proc ...entity_src_gen / pdbx_database_proc / pdbx_database_status / struct_biol
Item: _entity_src_gen.pdbx_gene_src_ncbi_taxonomy_id / _entity_src_gen.pdbx_gene_src_scientific_name / _pdbx_database_status.recvd_author_approval
Revision 1.3Dec 20, 2023Group: Data collection / Database references ...Data collection / Database references / Other / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_database_status / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_database_status.status_code_sf
Revision 1.4Nov 13, 2024Group: Structure summary / Category: pdbx_entry_details / pdbx_modification_feature / Item: _pdbx_entry_details.has_protein_modification

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: FAB HEAVY CHAIN
B: FAB LIGHT CHAIN
C: FAB HEAVY CHAIN
D: FAB LIGHT CHAIN
E: FAB HEAVY CHAIN
F: FAB LIGHT CHAIN
G: FAB HEAVY CHAIN
H: FAB HEAVY CHAIN
I: FAB LIGHT CHAIN
J: FAB HEAVY CHAIN
K: FAB LIGHT CHAIN
L: FAB LIGHT CHAIN
M: PROTEIN REV
N: PROTEIN REV
O: PROTEIN REV
P: PROTEIN REV
Q: PROTEIN REV
R: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)365,00318
Polymers365,00318
Non-polymers00
Water00
1
E: FAB HEAVY CHAIN
F: FAB LIGHT CHAIN
R: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5960 Å2
ΔGint-47.8 kcal/mol
Surface area27060 Å2
MethodPQS
2
C: FAB HEAVY CHAIN
D: FAB LIGHT CHAIN
P: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5910 Å2
ΔGint-47.3 kcal/mol
Surface area27030 Å2
MethodPQS
3
G: FAB HEAVY CHAIN
I: FAB LIGHT CHAIN
M: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5940 Å2
ΔGint-47.8 kcal/mol
Surface area26970 Å2
MethodPQS
4
J: FAB HEAVY CHAIN
K: FAB LIGHT CHAIN
Q: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5890 Å2
ΔGint-47.4 kcal/mol
Surface area27020 Å2
MethodPQS
5
H: FAB HEAVY CHAIN
L: FAB LIGHT CHAIN
N: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5880 Å2
ΔGint-47.7 kcal/mol
Surface area27030 Å2
MethodPQS
6
A: FAB HEAVY CHAIN
B: FAB LIGHT CHAIN
O: PROTEIN REV


Theoretical massNumber of molelcules
Total (without water)60,8343
Polymers60,8343
Non-polymers00
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5910 Å2
ΔGint-48.4 kcal/mol
Surface area27080 Å2
MethodPQS
Unit cell
Length a, b, c (Å)87.678, 87.692, 176.334
Angle α, β, γ (deg.)94.86, 95.50, 104.60
Int Tables number1
Space group name H-MP1

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Components

#1: Antibody
FAB HEAVY CHAIN


Mass: 24492.324 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21 / Variant (production host): CODONPLUSRIL
#2: Antibody
FAB LIGHT CHAIN


Mass: 23416.980 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others) / Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21 / Variant (production host): CODONPLUSRIL
#3: Protein
PROTEIN REV / ART/TRS / ANTI-REPRESSION TRANSACTIVATOR / REGULATOR OF EXPRESSION OF VIRAL PROTEINS


Mass: 12924.489 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 BH10
Production host: ESCHERICHIA COLI (E. coli) / Strain (production host): BL21 / Variant (production host): CODONPLUSRIL / References: UniProt: P04616
Has protein modificationY
Sequence detailsTHE C-TERMINAL DOMAIN OF REV IS DISORDERED IN THE CRYSTAL STRUCTURE. N-TERMINAL MET HAS BEEN ...THE C-TERMINAL DOMAIN OF REV IS DISORDERED IN THE CRYSTAL STRUCTURE. N-TERMINAL MET HAS BEEN CLEAVED OFF IN E. COLI AUTHOR PROVIDED GENBANK ACCESSION NUMBERS: CHAINS A,C,E,G,H,J GU223202 CHAINS B,D,F,I,K,L GU223201 CHAINS M,N,O,P,Q,R AAL29461

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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: 4.11 Å3/Da / Density % sol: 69.83 %
Description: 18 DIFFERENT MODELS USED IN PHASER AS AN ENSEMBLE
Crystal growpH: 8.5
Details: 12% POLYETHYLENE GLYCOL (PEG) 6000, 100 MM DI-AMMONIUM PHOSPHATE (DAP), AND 100 MM TRIS/HCL (PH 8.5)

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I02 / Wavelength: 0.97
DetectorType: ADSC CCD / Detector: CCD
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97 Å / Relative weight: 1
ReflectionResolution: 3.2→48.8 Å / Num. obs: 83664 / % possible obs: 99 % / Observed criterion σ(I): 0 / Redundancy: 3.4 % / Biso Wilson estimate: 77.96 Å2 / Rmerge(I) obs: 0.12 / Net I/σ(I): 9.9
Reflection shellResolution: 3.2→3.26 Å / Redundancy: 3.4 % / Rmerge(I) obs: 1 / Mean I/σ(I) obs: 1.1 / % possible all: 98.8

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Processing

Software
NameVersionClassification
BUSTER2.9.2refinement
HKL-2000data reduction
HKL-2000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRIES 1JPS, 2QR0, 2R8S, 3BDY, 1TZH, 1ZA3, 2R0K 2V7N, 3DVG, 1B2W, 1BEY, 1TZI, 2FJF, 2QQN, 1B4J, 2FGW, 2FJH, 3D85
Resolution: 3.17→48.78 Å / Cor.coef. Fo:Fc: 0.8909 / Cor.coef. Fo:Fc free: 0.892 / Cross valid method: THROUGHOUT / σ(F): 0
Details: IDEAL-DIST CONTACT TERM CONTACT SETUP. ALL ATOMS HAVE CCP4 ATOM TYPE FROM LIBRARY
RfactorNum. reflection% reflectionSelection details
Rfree0.25 4187 5.01 %RANDOM
Rwork0.2337 ---
obs0.2346 83636 --
Displacement parametersBiso mean: 143.62 Å2
Baniso -1Baniso -2Baniso -3
1--18.228 Å213.1177 Å2-6.3827 Å2
2---15.9372 Å2-4.1363 Å2
3---34.1652 Å2
Refine analyzeLuzzati coordinate error obs: 1.029 Å
Refinement stepCycle: LAST / Resolution: 3.17→48.78 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms22554 0 0 0 22554
Refine LS restraints
Refine-IDTypeDev idealNumberRestraint functionWeight
X-RAY DIFFRACTIONt_bond_d0.0123112HARMONIC2
X-RAY DIFFRACTIONt_angle_deg1.3231500HARMONIC2
X-RAY DIFFRACTIONt_dihedral_angle_d7632SINUSOIDAL2
X-RAY DIFFRACTIONt_incorr_chiral_ct
X-RAY DIFFRACTIONt_pseud_angle
X-RAY DIFFRACTIONt_trig_c_planes468HARMONIC2
X-RAY DIFFRACTIONt_gen_planes3378HARMONIC5
X-RAY DIFFRACTIONt_it23112HARMONIC20
X-RAY DIFFRACTIONt_nbd
X-RAY DIFFRACTIONt_omega_torsion2.69
X-RAY DIFFRACTIONt_other_torsion23.05
X-RAY DIFFRACTIONt_improper_torsion
X-RAY DIFFRACTIONt_chiral_improper_torsion3102SEMIHARMONIC5
X-RAY DIFFRACTIONt_sum_occupancies
X-RAY DIFFRACTIONt_utility_distance
X-RAY DIFFRACTIONt_utility_angle
X-RAY DIFFRACTIONt_utility_torsion
X-RAY DIFFRACTIONt_ideal_dist_contact25991SEMIHARMONIC4
LS refinement shellResolution: 3.17→3.25 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.2881 274 4.95 %
Rwork0.2777 5261 -
all0.2782 5535 -
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)
15.91580.04770.59166.8328-0.49649.63310.0923-1.27610.70960.3069-0.0723-0.8380.22951.0624-0.01990.67430.00050.0076-0.42830.071-0.51732.7648-0.197686.3652
21.15490.43350.08852.58170.90644.8127-0.0971-0.06120.09840.17530.1702-0.05690.19070.0117-0.07310.69310.0438-0.0254-0.68550.011-0.299734.3512-26.191341.5481
314.23824.866-1.462512.17670.7738.424-0.02180.96380.4171-0.35840.0507-1.0680.57221.4061-0.02890.7217-0.1240.0275-0.10620.0999-0.596148.7523-23.82465.7872
40.91880.3054-0.01553.7436-1.15584.5393-0.2384-0.0185-0.06160.16980.41290.14150.3271-0.361-0.17450.67960.03740.0467-0.6821-0.0011-0.354838.9756-42.707945.3623
56.54630.95590.15487.1662-0.63013.56230.0359-0.39930.37540.17450.0911.0642-0.9036-0.7306-0.12690.1182-0.34180.05060.4739-0.0238-0.5659-22.7754-31.107482.4664
62.99922.03890.32054.0363-0.6118.1101-0.09390.39130.2908-0.98980.04780.0449-0.54510.45440.04610.2952-0.22360.1019-0.17690.0752-0.3612-6.9363-26.06347.2141
76.6693-0.4252-0.25456.5831.45938.7290.2440.5656-1.0402-1.14120.00960.05021.01590.2241-0.2536-0.1904-0.3140.02220.44290.0522-0.4364-9.1771-54.373988.32
82.43871.2525-0.19462.79211.37134.451-0.22320.2531-0.3464-1.25140.192-0.3362-0.18531.17610.03120.6162-0.11380.2961-0.2491-0.0058-0.5389-10.2274-41.798240.4657
97.83332.45221.564616.3961-0.92684.3412-0.0691-0.6668-1.31610.7780.27510.62771.08090.5975-0.2060.0442-0.38630.18270.44720.0177-0.579817.9219-64.7313168.9562
102.6170.0690.3440.67890.31183.85260.15090.262-0.09890.0776-0.17780.07860.1708-0.01460.0269-0.4466-0.45050.02550.4807-0.1802-0.301416.4529-49.6849133.1572
113.51750.4651-1.35981.10490.01153.91810.2370.45740.27280.0084-0.09990.022-0.42110.3786-0.1371-0.4649-0.403-0.03750.4783-0.0272-0.324633.5699-49.9481129.3281
121.70060.9929-0.01054.0970.18378.83250.1489-0.823-0.05580.3519-0.27230.38990.9019-0.88370.1234-0.1563-0.47110.06450.21420.0849-0.51325.7662-9.5231127.452
131.1612-0.2836-0.53242.21030.10197.67990.0578-0.0454-0.1226-0.54120.0109-0.27330.0410.0201-0.06870.6550.1282-0.0745-0.6391-0.0015-0.037727.6102-3.412147.9094
141.0253-0.04520.54212.326-0.96743.38450.3336-0.9351-0.10490.1532-0.3452-0.30481.14750.02710.0116-0.056-0.413-0.0680.39170.1523-0.557320.125-2.3292134.217
152.7913-0.10940.42211.4138-0.79296.24420.0978-0.4454-0.15190.04560.0617-0.1846-0.0520.1863-0.1595-0.5834-0.18050.00870.424-0.1759-0.0539-7.3501-48.7364126.7643
162.8194-0.2679-0.96620.3792-0.20917.02590.102-0.68440.11840.10930.0764-0.0998-0.4447-0.0552-0.1784-0.6004-0.1612-0.02250.6317-0.1309-0.1904-24.4142-49.928130.5796
171.0308-0.9758-0.12693.0835-0.53936.04420.14120.0951-0.058-0.59540.04950.0828-0.4143-0.5462-0.19060.63910.1562-0.062-0.618-0.0191-0.168524.51913.400544.119
186.8838-0.68380.23868.2266-0.39173.43970.26290.2120.7679-0.5329-0.08950.5459-0.3599-1.1952-0.17350.6226-0.1685-0.01320.01390.0003-0.57716.72457.155192.1848
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1(CHAIN N)
2X-RAY DIFFRACTION2(CHAIN A)
3X-RAY DIFFRACTION3(CHAIN O)
4X-RAY DIFFRACTION4(CHAIN B)
5X-RAY DIFFRACTION5(CHAIN P)
6X-RAY DIFFRACTION6(CHAIN C)
7X-RAY DIFFRACTION7(CHAIN Q)
8X-RAY DIFFRACTION8(CHAIN D)
9X-RAY DIFFRACTION9(CHAIN R)
10X-RAY DIFFRACTION10(CHAIN E)
11X-RAY DIFFRACTION11(CHAIN F)
12X-RAY DIFFRACTION12(CHAIN G)
13X-RAY DIFFRACTION13(CHAIN H)
14X-RAY DIFFRACTION14(CHAIN I)
15X-RAY DIFFRACTION15(CHAIN J)
16X-RAY DIFFRACTION16(CHAIN K)
17X-RAY DIFFRACTION17(CHAIN L)
18X-RAY DIFFRACTION18(CHAIN M)

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