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- PDB-5mi0: A thermally stabilised version of Plasmodium falciparum RH5 -

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Basic information

Entry
Database: PDB / ID: 5mi0
TitleA thermally stabilised version of Plasmodium falciparum RH5
Components
  • (MONOCLONAL ANTIBODY 9AD4) x 2
  • Reticulocyte binding-like protein 5,Reticulocyte binding protein 5
KeywordsIMMUNE SYSTEM / malaria immunogen design Rosetta PROSS
Function / homology
Function and homology information


rhoptry lumen / rhoptry / symbiont entry into host / host cell membrane / bicellular tight junction / apical part of cell / heparin binding / cytoplasmic vesicle / host extracellular space / host cell surface receptor binding ...rhoptry lumen / rhoptry / symbiont entry into host / host cell membrane / bicellular tight junction / apical part of cell / heparin binding / cytoplasmic vesicle / host extracellular space / host cell surface receptor binding / host cell plasma membrane / protein-containing complex / extracellular region / membrane
Similarity search - Function
Rh5 coiled-coil domain / Rh5 coiled-coil domain / Immunoglobulins / Immunoglobulin-like / Sandwich / Mainly Beta
Similarity search - Domain/homology
Reticulocyte binding-like protein 5 / Reticulocyte binding protein 5 / Reticulocyte-binding protein homolog 5
Similarity search - Component
Biological speciesPlasmodium falciparum (malaria parasite P. falciparum)
Mus musculus (house mouse)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.35 Å
AuthorsCampeotto, I. / Goldenzweig, A. / Davey, J. / Barfod, L. / Marshall, J.M. / Silk, S.E. / Wright, K.E. / Draper, S.J. / Higgins, M.K. / Fleishman, S.J.
Funding support United Kingdom, 1items
OrganizationGrant numberCountry
United Kingdom
CitationJournal: Proc. Natl. Acad. Sci. U.S.A. / Year: 2017
Title: One-step design of a stable variant of the malaria invasion protein RH5 for use as a vaccine immunogen.
Authors: Campeotto, I. / Goldenzweig, A. / Davey, J. / Barfod, L. / Marshall, J.M. / Silk, S.E. / Wright, K.E. / Draper, S.J. / Higgins, M.K. / Fleishman, S.J.
History
DepositionNov 27, 2016Deposition site: PDBE / Processing site: PDBE
Revision 1.0Dec 28, 2016Provider: repository / Type: Initial release
Revision 1.1Jan 25, 2017Group: Database references
Revision 1.2Feb 8, 2017Group: Database references
Revision 1.3Jan 17, 2024Group: 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: Reticulocyte binding-like protein 5,Reticulocyte binding protein 5
B: MONOCLONAL ANTIBODY 9AD4
C: MONOCLONAL ANTIBODY 9AD4


Theoretical massNumber of molelcules
Total (without water)97,7433
Polymers97,7433
Non-polymers00
Water4,702261
1


  • Idetical with deposited unit
  • defined by software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5350 Å2
ΔGint-35 kcal/mol
Surface area33790 Å2
MethodPISA
Unit cell
Length a, b, c (Å)39.656, 85.547, 132.873
Angle α, β, γ (deg.)90.00, 90.91, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein Reticulocyte binding-like protein 5,Reticulocyte binding protein 5


Mass: 43842.387 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Plasmodium falciparum (malaria parasite P. falciparum)
Gene: Rh5 / Production host: Escherichia coli (E. coli)
References: UniProt: A0A159SJI5, UniProt: B2L3N7, UniProt: Q8IFM5*PLUS
#2: Antibody MONOCLONAL ANTIBODY 9AD4


Mass: 27641.205 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Oryctolagus cuniculus (rabbit)
#3: Antibody MONOCLONAL ANTIBODY 9AD4


Mass: 26259.277 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mus musculus (house mouse) / Production host: Oryctolagus cuniculus (rabbit)
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 261 / 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: 2.31 Å3/Da / Density % sol: 46.65 %
Crystal growTemperature: 277 K / Method: vapor diffusion, sitting drop
Details: 18% glycerol 22% PEG 1500 and additive: 0.02M HEPES sodium pH 6.8 and benzidine nicotinamide, pyromellitic acid and sulfaguanidine, each at 0.25% w/v

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I04-1 / Wavelength: 1 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Dec 4, 2015
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.35→42.77 Å / Num. obs: 36539 / % possible obs: 98.6 % / Redundancy: 6.5 % / Biso Wilson estimate: 49.1 Å2 / CC1/2: 0.996 / Rmerge(I) obs: 0.096 / Net I/σ(I): 13.9
Reflection shellResolution: 2.35→2.43 Å / Redundancy: 6.9 % / Rmerge(I) obs: 0.554 / Mean I/σ(I) obs: 3.1 / CC1/2: 0.832 / % possible all: 97.9

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Processing

Software
NameVersionClassification
BUSTER2.10.2refinement
PHENIX1.9refinement
XDSdata reduction
Aimlessdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4U0R
Resolution: 2.35→32.91 Å / Cor.coef. Fo:Fc: 0.9498 / Cor.coef. Fo:Fc free: 0.9201 / SU R Cruickshank DPI: 0.288 / Cross valid method: THROUGHOUT / σ(F): 0 / SU R Blow DPI: 0.284 / SU Rfree Blow DPI: 0.209 / SU Rfree Cruickshank DPI: 0.212
RfactorNum. reflection% reflectionSelection details
Rfree0.222 1798 4.93 %RANDOM
Rwork0.1763 ---
obs0.1786 36504 98.37 %-
Displacement parametersBiso mean: 50.44 Å2
Baniso -1Baniso -2Baniso -3
1-0.707 Å20 Å2-1.8019 Å2
2---2.1608 Å20 Å2
3---1.4538 Å2
Refine analyzeLuzzati coordinate error obs: 0.343 Å
Refinement stepCycle: 1 / Resolution: 2.35→32.91 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms5747 0 0 261 6008
Refine LS restraints
Refine-IDTypeDev idealNumberRestraint functionWeight
X-RAY DIFFRACTIONt_bond_d0.015891HARMONIC2
X-RAY DIFFRACTIONt_angle_deg1.117978HARMONIC2
X-RAY DIFFRACTIONt_dihedral_angle_d2023SINUSOIDAL2
X-RAY DIFFRACTIONt_incorr_chiral_ct
X-RAY DIFFRACTIONt_pseud_angle
X-RAY DIFFRACTIONt_trig_c_planes140HARMONIC2
X-RAY DIFFRACTIONt_gen_planes839HARMONIC5
X-RAY DIFFRACTIONt_it5891HARMONIC20
X-RAY DIFFRACTIONt_nbd
X-RAY DIFFRACTIONt_omega_torsion3.29
X-RAY DIFFRACTIONt_other_torsion18.9
X-RAY DIFFRACTIONt_improper_torsion
X-RAY DIFFRACTIONt_chiral_improper_torsion785SEMIHARMONIC5
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_contact6619SEMIHARMONIC4
LS refinement shellResolution: 2.35→2.42 Å / Total num. of bins used: 18
RfactorNum. reflection% reflection
Rfree0.3056 141 4.79 %
Rwork0.2156 2805 -
all0.2196 2946 -
obs--98.37 %
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)
10.11222.13650.13092.66681.893.2762-0.0033-0.01270.2016-0.1287-0.06220.02130.33150.05810.06550.286-0.0061-0.054-0.1469-0.1037-0.2384-9.2642-102.484477.097
24.03891.29241.77090.43090.84162.71670.26180.28270.08270.0466-0.2159-0.05471.0937-0.0557-0.04590.2462-0.02380.07-0.2587-0.0227-0.30643.7053-93.6417493.4601
30.27325.31141.35051.3737-1.61021.2730.03410.0528-0.2050.091-0.02610.2799-0.0228-0.1481-0.00790.5561-0.29320.0578-0.1926-0.1936-0.4623-15.8248-110.795479.6422
4-0.02571.10.80920.7436-3.06670.02570.0074-0.0119-0.0206-0.0731-0.02670.07240.03290.00540.01930.4083-0.0329-0.0724-0.1546-0.256-0.1781-21.5123-111.553465.337
53.07392.9409-1.43581.83710.13110.07430.0892-0.06740.0524-0.05870.01710.3266-0.0285-0.247-0.10630.084-0.27420.1256-0.1643-0.3084-0.2206-22.7598-104.507478.2258
62.03160.8827-0.77040.01070.65860.4360.0144-0.036-0.07390.06020.0956-0.00920.0711-0.1011-0.110.2691-0.28670.3057-0.0095-0.1818-0.1534-16.9197-99.0427493.6675
72.24211.89181.22082.20091.22523.59860.17430.17220.39140.2834-0.1787-0.01580.72080.1290.00440.04370.04910.0756-0.24960.0099-0.22928.3633-84.7508492.5039
81.68820.05650.19273.602-0.1440.00570.0233-0.18530.0139-0.267-0.03040.0652-0.19520.49770.00720.3754-0.1154-0.1255-0.10350.0316-0.4363-4.3168-90.785460.0685
9-0.05360.26011.67190-0.18761.95620.0019-0.00820.117-0.0674-0.04580.0308-0.0728-0.07480.04390.2880.1589-0.24990.0134-0.2103-0.1432-21.3619-101.86453.956
100.25321.30732.55011.0961-1.16491.191-0.003-0.04650.1910.0899-0.16870.2011-0.1416-0.01550.17180.1170.1557-0.2467-0.107-0.1994-0.1346-15.8158-95.7869465.5895
111.6090.88681.89621.92263.27744.4983-0.06620.17520.4556-0.4967-0.00430.3399-0.10140.11670.07050.0498-0.033-0.083-0.15560.0624-0.20525.5755-77.5755481.5053
122.51772.92264.48344.44365.41226.50240.08150.2715-0.062-0.388-0.05660.06340.04190.1191-0.02490.1753-0.0385-0.0358-0.04370.0005-0.1371-4.4141-94.3625471.9894
136.0190.2908-3.57162.6436-1.9127.1693-0.05850.08670.17850.12080.3561-0.4443-0.4023-0.4308-0.2976-0.14130.0291-0.0289-0.1772-0.0139-0.04192.7753-78.9322524.1342
145.71781.6160.81145.69743.040.054-0.15030.071-0.28040.24580.2601-0.0211-0.0819-0.165-0.1098-0.0918-0.06570.0239-0.0060.0466-0.0992-4.6731-90.7171526.9442
150.05530.23542.69743.9901-0.85815.2564-0.0153-0.0933-0.104-0.570.44330.23120.1328-0.9872-0.428-0.1261-0.1269-0.0640.11930.1148-0.0627-6.1365-82.7597514.8915
16-0.06820.41522.12440.19-4.44982.8126-0.0538-0.0280.2288-0.07870.1365-0.1264-0.4098-0.232-0.0827-0.1269-0.03920.034-0.1167-0.0160.04482.7325-74.4554518.0717
171.1946-2.4152-0.63454.8084-1.48423.7595-0.05460.03610.0935-0.55190.36490.04650.3112-0.8664-0.3103-0.037-0.1193-0.0213-0.0130.0075-0.0477-0.2935-87.8525520.244
180.44231.2348-1.58630-0.31863.71210.10360.10160.05720.01770.15770.1479-0.395-0.2067-0.2614-0.11420.2066-0.0028-0.08430.0678-0.0478-4.4685-82.0789541.6543
191.98411.1536-0.97262.21630.30950.0848-0.0648-0.11970.01770.29430.0940.14520.04750.1136-0.0292-0.16090.0487-0.008-0.02270.0085-0.0714-3.9135-96.9932568.2395
202.9198-4.0409-0.83784.44311.1453-0.02740.0095-0.4037-0.04560.18640.15360.21110.20920.3044-0.1631-0.18850.0221-0.0635-0.00070.0377-0.07221.9701-100.943566.2747
212.3082-0.1407-0.50760.00090.37335.21090.24230.21880.4066-0.0665-0.148-0.1812-0.462-0.0404-0.0942-0.10480.03590.0282-0.11150.03310.0260.6583-85.0593553.3903
220.32381.6649-2.92451.74521.29441.14060.01950.0875-0.284-0.2687-0.07850.0878-0.3134-0.15370.059-0.02380.0664-0.01410.01710.025-0.061-1.15-92.3642551.2943
233.8834-2.64551.58281.8035-1.49114.24530.0774-0.22170.37660.04350.0201-0.1322-0.2518-0.1455-0.0975-0.08480.0040.0137-0.0160.00590.0152.7127-89.069560.2377
246.49230.88450.42342.22373.02680.01180.155-0.26030.13150.27120.0029-0.2022-0.0243-0.079-0.158-0.14350.06660.01870.0822-0.0963-0.00330.0746-86.6503566.8697
251.47840.26731.64533.9394-3.55424.8136-0.3350.0963-0.3939-0.70060.40680.00821.0508-0.6012-0.07180.1579-0.24440.0816-0.1601-0.0478-0.20032.8808-109.281528.4704
262.3369-1.17471.93010.89180.4163.2823-0.04630.0519-0.0845-0.98850.1587-0.02310.7751-0.4004-0.11240.4794-0.2691-0.0478-0.24350.0101-0.23280.4846-103.248515.9468
270.55471.1205-1.59523.922-1.1173-0.0508-0.1064-0.1170.38320.04970.057-0.2931-0.046-0.00070.0494-0.0216-0.0070.0227-0.1821-0.0683-0.06878.8322-94.8462529.363
281.53-3.0555-1.3452.4796-4.23262.5632-0.09660.2579-0.0734-0.8178-0.0303-0.41330.83460.20970.1270.3618-0.11120.1962-0.2852-0.0238-0.072811.0554-103.946518.365
295.5826-0.87131.13544.13-4.87692.274-0.13740.0383-0.3297-0.53930.0056-0.25920.63860.22990.13180.2532-0.07370.1814-0.2247-0.0497-0.107810.7166-109.373526.0225
304.7397-0.56161.06316.3953-4.17121.7738-0.33380.0822-0.1342-0.66850.3719-0.04510.7004-0.1871-0.0381-0.0762-0.13510.0559-0.2842-0.0524-0.23513.4841-100.749524.7565
310.3136-0.8667-0.57893.1335-2.25960.3412-0.3790.2328-0.3684-0.45680.4881-0.01040.4411-0.1491-0.10910.0523-0.2232-0.03550.0545-0.0229-0.12410.4123-101.556527.6271
325.0116-2.5463-2.99372.28721.61531.96370.0856-0.2508-0.0793-0.14840.01520.03550.1197-0.0544-0.1007-0.21540.0152-0.0103-0.0677-0.0098-0.1047-5.1982-99.107559.58
33-0.01631.44110.7012.40991.5252.1875-0.06880.1083-0.26560.039-0.1890.46920.075-0.57950.2578-0.2163-0.0135-0.0290.0713-0.06410.012-10.566-107.501553.16
349.3502-3.7092-5.26942.38262.28082.83940.10250.4771-0.2799-0.1546-0.19150.1448-0.0171-0.45080.0891-0.19010.0118-0.03040.0304-0.0498-0.056-9.2519-100.658555.1861
350.54560.6449-0.00790.63740.29462.4605-0.03270.2206-0.22880.0418-0.1476-0.25550.0458-0.09170.18020.0549-0.03680.14160.0269-0.0230.1892-5.3566-115.214555.864
360.00261.74340.11510.0937-1.7292.899-0.0647-0.1547-0.0530.16660.076-0.06790.0128-0.1239-0.0113-0.11230.06670.0199-0.00360.05810.0004-13.7181-101.925570.7871
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1{ A|160 - A|174 }
2X-RAY DIFFRACTION2{ A|175 - A|223 }
3X-RAY DIFFRACTION3{ A|224 - A|235 }
4X-RAY DIFFRACTION4{ A|236 - A|302 }
5X-RAY DIFFRACTION5{ A|303 - A|318 }
6X-RAY DIFFRACTION6{ A|319 - A|324 }
7X-RAY DIFFRACTION7{ A|325 - A|373 }
8X-RAY DIFFRACTION8{ A|374 - A|390 }
9X-RAY DIFFRACTION9{ A|391 - A|409 }
10X-RAY DIFFRACTION10{ A|410 - A|423 }
11X-RAY DIFFRACTION11{ A|424 - A|453 }
12X-RAY DIFFRACTION12{ A|454 - A|501 }
13X-RAY DIFFRACTION13{ B|2 - B|36 }
14X-RAY DIFFRACTION14{ B|37 - B|49 }
15X-RAY DIFFRACTION15{ B|50 - B|70 }
16X-RAY DIFFRACTION16{ B|71 - B|82 }
17X-RAY DIFFRACTION17{ B|83 - B|110 }
18X-RAY DIFFRACTION18{ B|111 - B|131 }
19X-RAY DIFFRACTION19{ B|132 - B|138 }
20X-RAY DIFFRACTION20{ B|139 - B|150 }
21X-RAY DIFFRACTION21{ B|151 - B|167 }
22X-RAY DIFFRACTION22{ B|168 - B|182 }
23X-RAY DIFFRACTION23{ B|183 - B|213 }
24X-RAY DIFFRACTION24{ B|214 - B|222 }
25X-RAY DIFFRACTION25{ C|1 - C|20 }
26X-RAY DIFFRACTION26{ C|21 - C|42 }
27X-RAY DIFFRACTION27{ C|43 - C|51 }
28X-RAY DIFFRACTION28{ C|52 - C|73 }
29X-RAY DIFFRACTION29{ C|74 - C|84 }
30X-RAY DIFFRACTION30{ C|85 - C|97 }
31X-RAY DIFFRACTION31{ C|98 - C|111 }
32X-RAY DIFFRACTION32{ C|112 - C|142 }
33X-RAY DIFFRACTION33{ C|143 - C|159 }
34X-RAY DIFFRACTION34{ C|160 - C|202 }
35X-RAY DIFFRACTION35{ C|203 - C|211 }
36X-RAY DIFFRACTION36{ C|212 - C|217 }

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