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Basic information

Entry
Database: PDB / ID: 4n9g
TitleCrystal Structure of a Computationally Designed RSV-Presenting Epitope Scaffold And Its Elicited Antibody 17HD9
Components
  • Antibody 17HD9, Heavy Chain
  • Antibody 17HD9, Light Chain
  • Epitope Scaffold rsv_1isea_FFL_001_C
KeywordsIMMUNE SYSTEM / Antibody / Hairpin / Epitope Scaffold
Function / homologyImmunoglobulins / Immunoglobulin-like / Sandwich / Mainly Beta
Function and homology information
Biological speciesMacaca mulatta (Rhesus monkey)
synthetic construct (others)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.5 Å
AuthorsCarrico, C.T.D. / Strong, R.K.
CitationJournal: Nature / Year: 2014
Title: Proof of principle for epitope-focused vaccine design.
Authors: Correia, B.E. / Bates, J.T. / Loomis, R.J. / Baneyx, G. / Carrico, C. / Jardine, J.G. / Rupert, P. / Correnti, C. / Kalyuzhniy, O. / Vittal, V. / Connell, M.J. / Stevens, E. / Schroeter, A. ...Authors: Correia, B.E. / Bates, J.T. / Loomis, R.J. / Baneyx, G. / Carrico, C. / Jardine, J.G. / Rupert, P. / Correnti, C. / Kalyuzhniy, O. / Vittal, V. / Connell, M.J. / Stevens, E. / Schroeter, A. / Chen, M. / Macpherson, S. / Serra, A.M. / Adachi, Y. / Holmes, M.A. / Li, Y. / Klevit, R.E. / Graham, B.S. / Wyatt, R.T. / Baker, D. / Strong, R.K. / Crowe, J.E. / Johnson, P.R. / Schief, W.R.
History
DepositionOct 21, 2013Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 12, 2014Provider: repository / Type: Initial release
Revision 1.1Sep 20, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Antibody 17HD9, Heavy Chain
B: Antibody 17HD9, Light Chain
C: Epitope Scaffold rsv_1isea_FFL_001_C
D: Epitope Scaffold rsv_1isea_FFL_001_C
E: Antibody 17HD9, Heavy Chain
F: Antibody 17HD9, Light Chain
H: Antibody 17HD9, Heavy Chain
L: Antibody 17HD9, Light Chain
M: Antibody 17HD9, Heavy Chain
N: Antibody 17HD9, Light Chain
Y: Epitope Scaffold rsv_1isea_FFL_001_C
Z: Epitope Scaffold rsv_1isea_FFL_001_C


Theoretical massNumber of molelcules
Total (without water)249,70012
Polymers249,70012
Non-polymers00
Water1,06359
1
A: Antibody 17HD9, Heavy Chain
B: Antibody 17HD9, Light Chain
C: Epitope Scaffold rsv_1isea_FFL_001_C


Theoretical massNumber of molelcules
Total (without water)62,4253
Polymers62,4253
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
D: Epitope Scaffold rsv_1isea_FFL_001_C
E: Antibody 17HD9, Heavy Chain
F: Antibody 17HD9, Light Chain


Theoretical massNumber of molelcules
Total (without water)62,4253
Polymers62,4253
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
H: Antibody 17HD9, Heavy Chain
L: Antibody 17HD9, Light Chain
Y: Epitope Scaffold rsv_1isea_FFL_001_C


Theoretical massNumber of molelcules
Total (without water)62,4253
Polymers62,4253
Non-polymers00
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
M: Antibody 17HD9, Heavy Chain
N: Antibody 17HD9, Light Chain
Z: Epitope Scaffold rsv_1isea_FFL_001_C


Theoretical massNumber of molelcules
Total (without water)62,4253
Polymers62,4253
Non-polymers00
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)64.213, 89.266, 104.304
Angle α, β, γ (deg.)89.99, 102.73, 89.91
Int Tables number1
Space group name H-MP1
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11H
21M
12L
22N
13A
23E
14B
24F
15C
25D
16Y
26Z

NCS domain segments:
Dom-IDComponent-IDEns-IDRefine codeAuth asym-IDAuth seq-ID
1114H1 - 999
2114M1 - 999
1124L1 - 999
2124N1 - 999
1134A1 - 999
2134E1 - 999
1144B1 - 999
2144F1 - 999
1154C1 - 999
2154D1 - 999
1164Y1 - 999
2164Z1 - 999

NCS ensembles :
ID
1
2
3
4
5
6

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Components

#1: Antibody
Antibody 17HD9, Heavy Chain


Mass: 24461.527 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Macaca mulatta (Rhesus monkey) / Plasmid: pHLsec / Cell line (production host): HEK 293F / Production host: Homo sapiens (human)
#2: Antibody
Antibody 17HD9, Light Chain


Mass: 23512.922 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Macaca mulatta (Rhesus monkey) / Plasmid: pHLsec / Cell line (production host): HEK 293F / Production host: Homo sapiens (human)
#3: Protein
Epitope Scaffold rsv_1isea_FFL_001_C


Mass: 14450.532 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) synthetic construct (others)
Gene: Computationally designed variant of Ribosomal Recycling Factor
Plasmid: pET29b / Production host: Escherichia coli (E. coli) / Strain (production host): BL21
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 59 / 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: 2.34 Å3/Da / Density % sol: 47.33 %
Crystal growTemperature: 298 K / Method: vapor diffusion, hanging drop / pH: 5.6
Details: 14% PEG4000, 0.1M sodium acetate, 0.2M ammonium sulfate, pH 5.6, VAPOR DIFFUSION, HANGING DROP, temperature 298K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 5.0.2 / Wavelength: 1 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Aug 18, 2013
RadiationMonochromator: Double-crystal, Si(111) liquid N2 cooled / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.5→48.75 Å / Num. all: 78170 / Num. obs: 76354 / % possible obs: 97.7 % / Observed criterion σ(F): 5 / Observed criterion σ(I): 5 / Redundancy: 3.9 % / Rmerge(I) obs: 0.076 / Net I/σ(I): 9.4
Reflection shellResolution: 2.5→2.59 Å / Redundancy: 3.89 % / Rmerge(I) obs: 0.212 / Mean I/σ(I) obs: 4.4 / % possible all: 97.5

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Processing

Software
NameVersionClassification
REFMAC5.5.0109refinement
d*TREKdata reduction
d*TREKdata scaling
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 17HD9 N TERMINAL DOMAIN PAIR: PDB ENTRY 3Q6G AND 3EO9. 17HD9 C TERMINAL DOMAIN PAIR: PDB ENTRY 3QHZ AND 3SOB. SCAFFOLD: UNPUBLISHED CRYSTAL STRUCTURE OF UNBOUND SCAFFOLD ALL MODELS ...Starting model: 17HD9 N TERMINAL DOMAIN PAIR: PDB ENTRY 3Q6G AND 3EO9. 17HD9 C TERMINAL DOMAIN PAIR: PDB ENTRY 3QHZ AND 3SOB. SCAFFOLD: UNPUBLISHED CRYSTAL STRUCTURE OF UNBOUND SCAFFOLD ALL MODELS HAD EXPECTED CONTACT RESIDUES REMOVED
Resolution: 2.5→48.75 Å / Cor.coef. Fo:Fc: 0.864 / Cor.coef. Fo:Fc free: 0.829 / SU B: 23.784 / SU ML: 0.255 / Cross valid method: THROUGHOUT / ESU R: 0.649 / ESU R Free: 0.336 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.29516 3829 5 %RANDOM
Rwork0.26335 ---
all0.26496 76354 --
obs0.26496 72525 97.68 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 41.108 Å2
Baniso -1Baniso -2Baniso -3
1-0.14 Å2-0.04 Å2-0.02 Å2
2---0.02 Å2-0.05 Å2
3----0.13 Å2
Refinement stepCycle: LAST / Resolution: 2.5→48.75 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14202 0 0 59 14261
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0050.02214537
X-RAY DIFFRACTIONr_bond_other_d0.0010.029526
X-RAY DIFFRACTIONr_angle_refined_deg0.9611.95519805
X-RAY DIFFRACTIONr_angle_other_deg0.74323409
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.83651885
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.87324.908546
X-RAY DIFFRACTIONr_dihedral_angle_3_deg13.612152280
X-RAY DIFFRACTIONr_dihedral_angle_4_deg15.5631547
X-RAY DIFFRACTIONr_chiral_restr0.0550.22277
X-RAY DIFFRACTIONr_gen_planes_refined0.0030.02116209
X-RAY DIFFRACTIONr_gen_planes_other0.0010.022762
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_it0.33229426
X-RAY DIFFRACTIONr_mcbond_other0.04823801
X-RAY DIFFRACTIONr_mcangle_it0.622315241
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it0.75445111
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it1.19264557
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

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

Ens-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1H2703MEDIUM POSITIONAL0.260.5
1H2703MEDIUM THERMAL0.122
2L2704MEDIUM POSITIONAL0.240.5
2L2704MEDIUM THERMAL0.142
3A2666MEDIUM POSITIONAL0.280.5
3A2666MEDIUM THERMAL0.122
4B2687MEDIUM POSITIONAL0.270.5
4B2687MEDIUM THERMAL0.132
5C414MEDIUM POSITIONAL0.40.5
5C414MEDIUM THERMAL0.142
6Y435MEDIUM POSITIONAL0.380.5
6Y435MEDIUM THERMAL0.192
LS refinement shellResolution: 2.5→2.565 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.4 264 -
Rwork0.315 5430 -
obs-5430 97.53 %
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)
12.8210.59041.95862.6883-0.71263.2693-0.2809-0.06250.1182-0.0613-0.10920.2525-0.43-0.09210.39010.1862-0.0138-0.04320.044-0.02340.164926.1007-62.01534.3363
22.91691.97370.78614.53470.17941.4705-0.09340.07830.0041-0.09140.11250.1322-0.04990.0264-0.01910.03950.0137-0.02120.15150.00460.08978.3516-80.94459.6627
32.24150.79570.72762.5896-1.22762.35020.00730.11920.0811-0.05590.00030.01750.08790.2476-0.00760.06090.04160.01730.10890.01070.089832.8675-82.008337.2271
41.30720.068-0.29782.1504-0.7194.68970.01540.042-0.0696-0.0081-0.0393-0.06920.26330.02060.02390.0367-0.0185-0.03870.0909-0.02280.118720.4335-89.53153.9013
52.63961.6221-2.71361.1196-0.104721.10590.0533-0.0649-0.21050.0354-0.0333-0.0896-0.87630.7706-0.02010.2833-0.2086-0.03860.17140.05130.140741.9365-59.487154.7514
64.7553-2.3281-0.35851.4183-3.194623.4453-0.1151-0.06080.0680.13330.0237-0.0406-1.1958-0.65510.09140.37450.2495-0.06550.147-0.05650.07367.3754-14.3931-59.1563
71.9814-0.51321.56712.15561.05593.6825-0.2481-0.00490.08440.0175-0.0765-0.2143-0.51290.01860.32460.21560.046-0.05150.05910.01730.164622.5266-17.3352-39.6386
81.924-0.83390.93573.324-0.49282.0773-0.0895-0.08610.057-0.04580.0792-0.131-0.05010.03430.01040.05870.0165-0.0060.098-0.0080.099239.8667-35.3809-15.8808
92.3696-1.05341.11022.83520.99882.11520.0528-0.10220.0268-0.0082-0.11120.09360.0845-0.2290.05830.0605-0.00030.0130.1148-0.01250.105116.0001-37.5246-42.4181
101.1771-0.6431-0.58882.19140.29913.6087-0.0352-0.0341-0.10260.1159-0.04340.13190.2218-0.10360.07870.0320.0142-0.01890.11320.01050.119128.3003-44.9001-9.1534
110.7628-0.1211-0.99461.7514-1.22934.61760.06470.038-0.029-0.1058-0.01070.07530.3359-0.1637-0.0540.11230.0327-0.01310.0730.00950.110453.8833-95.5081-34.1935
121.6522-0.32660.00823.74680.39482.52880.0548-0.14590.1318-0.0735-0.08370.2587-0.0497-0.16120.02890.00520.00750.01760.12730.00370.152839.858-79.4286-7.2331
130.2222-0.1798-0.41530.35470.42082.5550.0425-0.03420.0178-0.0585-0.0431-0.1105-0.13910.12620.00060.06260.00680.02050.0790.01940.168258.8681-74.3924-26.1642
140.7174-0.53321.03951.559-0.55074.6687-0.06190.00230.04340.09230.008-0.0475-0.1150.18260.05390.0777-0.00140.00930.0906-0.00910.142853.2825-70.94231.3117
151.05780.2897-0.86991.90231.90185.04020.0732-0.0187-0.03330.17230.0167-0.10690.27520.2156-0.08990.1056-0.0212-0.01840.0791-0.00460.130259.011-51.036128.9308
161.85220.4558-0.4983.4351-1.00332.85750.04410.10990.1152-0.0209-0.0555-0.224-0.03170.14880.01140.01590.0250.0230.12280.00650.130673.147-34.7252.1241
171.26880.5009-0.78132.7866-0.46161.75140.04180.04540.04880.0453-0.03070.1637-0.1187-0.1042-0.01120.06640.01490.03170.117-0.00170.137251.1433-31.165430.541
181.32740.20460.81461.66810.60694.4335-0.05380.02980.0447-0.05850.00030.1166-0.1415-0.1160.05350.0495-0.00390.01990.0670.0240.130660.6625-26.1907-4.8155
190.27870.85761.30633.03761.155815.53380.0089-0.04030.0685-0.3092-0.1016-0.00980.38960.45110.09270.2650.23280.02510.2114-0.03980.160965.7073-95.752-56.1328
203.48681.15530.94566.36220.56787.85960.31360.01910.13280.4327-0.49860.06250.4735-0.38290.1850.2427-0.20090.03230.193-0.00880.085347.4832-51.61348.392
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A1 - 109
2X-RAY DIFFRACTION2A110 - 216
3X-RAY DIFFRACTION3B1 - 106
4X-RAY DIFFRACTION4B107 - 214
5X-RAY DIFFRACTION5C63 - 97
6X-RAY DIFFRACTION6D62 - 95
7X-RAY DIFFRACTION7E1 - 104
8X-RAY DIFFRACTION8E105 - 213
9X-RAY DIFFRACTION9F2 - 106
10X-RAY DIFFRACTION10F107 - 213
11X-RAY DIFFRACTION11H1 - 113
12X-RAY DIFFRACTION12H114 - 214
13X-RAY DIFFRACTION13L1 - 149
14X-RAY DIFFRACTION14L150 - 214
15X-RAY DIFFRACTION15M2 - 125
16X-RAY DIFFRACTION16M126 - 225
17X-RAY DIFFRACTION17N1 - 107
18X-RAY DIFFRACTION18N108 - 213
19X-RAY DIFFRACTION19Y63 - 95
20X-RAY DIFFRACTION20Z62 - 87

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