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- PDB-6bi2: Trastuzumab Fab D185A (Light Chain) Mutant Biotin Conjugation. -

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Basic information

Entry
Database: PDB / ID: 6bi2
TitleTrastuzumab Fab D185A (Light Chain) Mutant Biotin Conjugation.
Components
  • Trastuzumab Anti-HER2 Fab Heavy Chain
  • Trastuzumab Anti-HER2 Fab Light Chain
KeywordsIMMUNE SYSTEM / Fab / immunoglobulin
Function / homologyImmunoglobulins / Immunoglobulin-like / Sandwich / Mainly Beta / BIOTIN
Function and homology information
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.801 Å
AuthorsDiDonato, M. / Spraggon, G.
CitationJournal: Chembiochem / Year: 2018
Title: Tuning a Protein-Labeling Reaction to Achieve Highly Site Selective Lysine Conjugation.
Authors: Pham, G.H. / Ou, W. / Bursulaya, B. / DiDonato, M. / Herath, A. / Jin, Y. / Hao, X. / Loren, J. / Spraggon, G. / Brock, A. / Uno, T. / Geierstanger, B.H. / Cellitti, S.E.
History
DepositionOct 31, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0Feb 14, 2018Provider: repository / Type: Initial release
Revision 1.1Mar 28, 2018Group: Data collection / Database references / Category: citation / citation_author / Item: _citation.title / _citation_author.name
Revision 1.2Apr 25, 2018Group: Data collection / Database references / Category: citation
Item: _citation.journal_volume / _citation.page_first / _citation.page_last
Revision 1.3Oct 4, 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
H: Trastuzumab Anti-HER2 Fab Heavy Chain
L: Trastuzumab Anti-HER2 Fab Light Chain
I: Trastuzumab Anti-HER2 Fab Heavy Chain
M: Trastuzumab Anti-HER2 Fab Light Chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)95,74513
Polymers94,8224
Non-polymers9239
Water13,367742
1
H: Trastuzumab Anti-HER2 Fab Heavy Chain
L: Trastuzumab Anti-HER2 Fab Light Chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)47,9037
Polymers47,4112
Non-polymers4935
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4640 Å2
ΔGint-6 kcal/mol
Surface area19730 Å2
MethodPISA
2
I: Trastuzumab Anti-HER2 Fab Heavy Chain
M: Trastuzumab Anti-HER2 Fab Light Chain
hetero molecules


Theoretical massNumber of molelcules
Total (without water)47,8416
Polymers47,4112
Non-polymers4314
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4270 Å2
ΔGint-9 kcal/mol
Surface area19170 Å2
MethodPISA
Unit cell
Length a, b, c (Å)38.646, 69.139, 91.471
Angle α, β, γ (deg.)109.80, 100.29, 90.08
Int Tables number1
Space group name H-MP1

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Components

#1: Antibody Trastuzumab Anti-HER2 Fab Heavy Chain


Mass: 23988.850 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Cell line (production host): HEK293 / Production host: Homo sapiens (human)
#2: Antibody Trastuzumab Anti-HER2 Fab Light Chain


Mass: 23422.021 Da / Num. of mol.: 2 / Mutation: D185A
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Cell line (production host): HEK293 / Production host: Homo sapiens (human)
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 7 / Source method: obtained synthetically / Formula: C2H6O2
#4: Chemical ChemComp-BTN / BIOTIN / Biotin


Mass: 244.311 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C10H16N2O3S / Feature type: SUBJECT OF INVESTIGATION
#5: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 742 / 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: 2.38 Å3/Da / Density % sol: 48.34 %
Crystal growTemperature: 294 K / Method: vapor diffusion, sitting drop / pH: 7 / Details: 1.0% trptone, 20% PEG 3350, 100 mM HEPES pH 7.0

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 5.0.3 / Wavelength: 0.97648 Å
DetectorType: ADSC QUANTUM 315r / Detector: CCD / Date: Jul 13, 2017
RadiationMonochromator: Single crystal, cylindrically bent, Si(220) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97648 Å / Relative weight: 1
ReflectionResolution: 1.8→50 Å / Num. obs: 78712 / % possible obs: 97.5 % / Redundancy: 3.9 % / Biso Wilson estimate: 29.76 Å2 / Rmerge(I) obs: 0.047 / Rpim(I) all: 0.028 / Rrim(I) all: 0.055 / Χ2: 0.874 / Net I/σ(I): 25
Reflection shellResolution: 1.8→1.86 Å / Redundancy: 3.7 % / Rmerge(I) obs: 0.461 / Mean I/σ(I) obs: 2.14 / Num. unique obs: 7683 / CC1/2: 0.882 / Rpim(I) all: 0.276 / Rrim(I) all: 0.538 / Χ2: 0.599 / % possible all: 95.1

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Processing

Software
NameVersionClassification
PHENIX(1.12_2829: ???)refinement
HKL-2000data reduction
SCALEPACKdata scaling
PHASERphasing
Cootmodel building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 1N8Z
Resolution: 1.801→37.37 Å / SU ML: 0.2 / Cross valid method: FREE R-VALUE / σ(F): 1.99 / Phase error: 21.99
RfactorNum. reflection% reflection
Rfree0.2021 3780 4.8 %
Rwork0.1658 --
obs0.1675 78687 97.28 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso mean: 42.1 Å2
Refinement stepCycle: LAST / Resolution: 1.801→37.37 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6551 0 58 742 7351
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0076981
X-RAY DIFFRACTIONf_angle_d0.9089530
X-RAY DIFFRACTIONf_dihedral_angle_d12.4464183
X-RAY DIFFRACTIONf_chiral_restr0.0591066
X-RAY DIFFRACTIONf_plane_restr0.0061226
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.8015-1.82430.2631470.25582557X-RAY DIFFRACTION90
1.8243-1.84830.2511290.2462723X-RAY DIFFRACTION95
1.8483-1.87360.28241650.23122681X-RAY DIFFRACTION96
1.8736-1.90040.30671290.22382758X-RAY DIFFRACTION97
1.9004-1.92870.26951380.2132792X-RAY DIFFRACTION96
1.9287-1.95890.24031200.212737X-RAY DIFFRACTION97
1.9589-1.9910.20921380.20722754X-RAY DIFFRACTION96
1.991-2.02530.30271410.20242818X-RAY DIFFRACTION97
2.0253-2.06210.23621510.19342696X-RAY DIFFRACTION97
2.0621-2.10180.22431450.19132755X-RAY DIFFRACTION97
2.1018-2.14470.23221500.192813X-RAY DIFFRACTION97
2.1447-2.19130.25471580.18242723X-RAY DIFFRACTION98
2.1913-2.24230.23371560.18222772X-RAY DIFFRACTION98
2.2423-2.29830.26721540.18082779X-RAY DIFFRACTION98
2.2983-2.36050.23731390.18782761X-RAY DIFFRACTION97
2.3605-2.42990.2551250.18692818X-RAY DIFFRACTION98
2.4299-2.50830.21991340.18542791X-RAY DIFFRACTION98
2.5083-2.5980.20631490.18272798X-RAY DIFFRACTION98
2.598-2.7020.23791440.18172795X-RAY DIFFRACTION98
2.702-2.82490.21541310.17542803X-RAY DIFFRACTION98
2.8249-2.97380.19811410.17412836X-RAY DIFFRACTION99
2.9738-3.160.20451310.17222800X-RAY DIFFRACTION98
3.16-3.40380.18361470.1662804X-RAY DIFFRACTION99
3.4038-3.74610.21521360.15272824X-RAY DIFFRACTION99
3.7461-4.28750.17091110.14022878X-RAY DIFFRACTION99
4.2875-5.39920.11971190.11962846X-RAY DIFFRACTION99
5.3992-37.37840.16131520.14362795X-RAY DIFFRACTION98
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)
13.27143.2857-0.8737.5659-2.81084.036-0.0447-0.10860.09290.20320.0350.0677-0.36550.16810.04510.2295-0.00680.01140.2663-0.01480.235816.65431.8558-6.7471
21.50410.29170.09423.1975-0.08153.6795-0.0717-0.02310.1464-0.08910.07250.4183-0.2792-0.17270.00430.17840.01260.0190.17940.00280.28479.397132.001-11.539
32.7566-0.8897-0.87144.02480.4822.6732-0.03440.3658-0.269-0.07150.0946-0.1740.49520.29040.02240.29770.0014-0.05740.2929-0.0850.205326.56662.2451-14.7945
44.4438-1.0646-2.1953.26721.79184.70030.0453-0.0614-0.04970.13290.2297-0.420.00430.5714-0.26130.25990.0089-0.03240.2266-0.06980.260531.90134.1616-16.2061
58.884-4.7188-2.06924.67051.90432.07190.09050.3818-0.2244-0.2046-0.20780.22660.16-0.40090.07220.3312-0.0279-0.03970.30830.00180.195210.033226.4089-36.5598
63.03640.9734-0.88583.6072-0.09665.47110.03220.30570.4448-0.2297-0.1025-0.113-0.4106-0.07560.08090.23670.02590.02370.20590.01590.203514.073534.8986-31.693
73.0611-2.803-3.67744.97432.15248.6916-0.08460.13840.05-0.0278-0.1010.1641-0.0879-0.17110.13980.2157-0.0276-0.05070.2379-0.0010.186112.628429.2733-29.2456
89.3296-8.6328-1.51528.7662.15441.04410.10190.4532-0.0866-0.2275-0.3696-0.03290.33690.29370.30360.58690.08750.09950.3250.01690.268123.397113.8426-38.7984
94.96451.0914-2.3555.1977-4.12354.06090.2226-0.4221-0.29130.9061-0.1737-0.2571-0.38920.7081-0.14720.5620.0756-0.07830.2466-0.06530.362229.4805-8.4038-17.7125
102.30910.8148-1.17728.236-6.26775.73730.35390.3954-0.0962-0.40590.10250.586-0.1911-0.1596-0.44660.45650.1067-0.02470.3231-0.07960.31423.2542-1.2937-30.8956
111.3518-1.0683-0.18869.0211-0.9684.00360.33830.2597-0.5756-0.591-0.39760.71540.1551-0.0041-0.10630.35880.0363-0.12980.3184-0.12780.473617.5419-6.3159-27.1873
123.0614-1.71220.68222.098-2.43319.25210.23910.23780.3269-0.1612-0.1741-0.5934-1.67230.92490.09460.6597-0.06470.08080.4359-0.00780.349428.167813.3526-32.3872
131.7763-0.5529-0.26069.1754-2.04324.28910.20280.1611-0.4185-0.2332-0.16080.64540.49210.01750.01090.35290.0092-0.07350.2562-0.0760.394722.6991-11.7486-24.5253
142.25191.183-0.65420.7615-1.18658.02890.23120.3369-0.4566-1.7121-0.2276-0.09740.60640.2036-0.06710.75930.1514-0.06110.3172-0.12470.327227.8934-8.9779-34.6806
156.1498-4.9165-0.5258.54091.49043.92350.0570.2185-0.3224-0.1513-0.26490.4421-0.2257-0.1380.22540.1801-0.07430.02620.2091-0.01390.184324.846825.07566.3158
168.4771-3.4779-0.8386.19331.24276.62160.0361-0.06140.44870.1798-0.10280.1597-0.85260.05410.00610.2635-0.01430.00490.20330.0180.187224.920936.34996.1008
173.650.2171-0.32596.34150.49553.82910.076-0.09770.28530.3006-0.0998-0.6082-0.38030.54160.05640.2536-0.1059-0.03860.275-0.02270.263935.435331.857212.2152
180.63611.5211-0.3613.646-1.16982.9137-0.0591-0.01690.0738-0.084-0.2031-0.6451-0.16560.3460.2830.2004-0.0490.03620.21380.00060.247732.252228.94674.2109
195.1361-1.644-1.18843.31333.85024.7647-0.1027-0.25650.6040.415-0.08820.0608-0.88270.07730.17080.5177-0.0582-0.04570.2382-0.05840.260626.378935.084516.9681
201.4768-0.02640.77012.4658-1.59893.40950.0444-0.3504-0.2336-0.07390.43050.3910.2831-0.9443-0.24050.2433-0.0679-0.01060.42910.12930.279612.55735.169315.1155
217.09576.0518-5.44786.7139-4.32334.25590.0134-0.1274-0.6715-0.4010.49040.31710.5297-1.8408-0.34410.2978-0.3185-0.14370.77560.26830.43795.58371.48039.8747
222.2699-0.7189-0.83492.895-0.13276.3428-0.0544-0.62220.29340.99110.03970.0326-0.18430.6590.030.7401-0.0259-0.03080.513-0.10510.282327.537631.01233.0122
231.13780.7137-0.00483.9571-2.28963.28550.0123-0.1782-0.06480.410.29750.0224-0.1467-0.1847-0.26080.21220.0903-0.00020.42450.01380.253321.41110.435526.326
241.9706-0.58010.46914.1984-0.89165.80520.111-0.2909-0.3363-0.30990.35740.08480.3332-0.6323-0.3140.2799-0.0995-0.05050.40780.14060.285918.0483-4.684429.0089
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'H' and (resid 1 through 32 )
2X-RAY DIFFRACTION2chain 'H' and (resid 33 through 113 )
3X-RAY DIFFRACTION3chain 'H' and (resid 114 through 141 )
4X-RAY DIFFRACTION4chain 'H' and (resid 142 through 222 )
5X-RAY DIFFRACTION5chain 'L' and (resid 1 through 25 )
6X-RAY DIFFRACTION6chain 'L' and (resid 26 through 75 )
7X-RAY DIFFRACTION7chain 'L' and (resid 76 through 101 )
8X-RAY DIFFRACTION8chain 'L' and (resid 102 through 113 )
9X-RAY DIFFRACTION9chain 'L' and (resid 114 through 128 )
10X-RAY DIFFRACTION10chain 'L' and (resid 129 through 150 )
11X-RAY DIFFRACTION11chain 'L' and (resid 151 through 163 )
12X-RAY DIFFRACTION12chain 'L' and (resid 164 through 174 )
13X-RAY DIFFRACTION13chain 'L' and (resid 175 through 197 )
14X-RAY DIFFRACTION14chain 'L' and (resid 198 through 213 )
15X-RAY DIFFRACTION15chain 'I' and (resid 1 through 17 )
16X-RAY DIFFRACTION16chain 'I' and (resid 18 through 32 )
17X-RAY DIFFRACTION17chain 'I' and (resid 33 through 67 )
18X-RAY DIFFRACTION18chain 'I' and (resid 68 through 91 )
19X-RAY DIFFRACTION19chain 'I' and (resid 92 through 113 )
20X-RAY DIFFRACTION20chain 'I' and (resid 114 through 210 )
21X-RAY DIFFRACTION21chain 'I' and (resid 211 through 220 )
22X-RAY DIFFRACTION22chain 'M' and (resid 1 through 90 )
23X-RAY DIFFRACTION23chain 'M' and (resid 91 through 128 )
24X-RAY DIFFRACTION24chain 'M' and (resid 129 through 214 )

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