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- PDB-13cv: Crystal structure of covalent inhibitor 3-(2-chloroacetamido)-N-(... -

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Basic information

Entry
Database: PDB / ID: 13cv
TitleCrystal structure of covalent inhibitor 3-(2-chloroacetamido)-N-(pyridin-2-yl)benzamide bound to Ubiquitin C-terminal Hydrolase-L3
ComponentsUbiquitin carboxyl-terminal hydrolase isozyme L3
KeywordsHYDROLASE/HYDROLASE INHIBITOR / UCHL3 / covalent inhibitor / chloroacetamide / HYDROLASE / HYDROLASE-HYDROLASE INHIBITOR complex
Function / homology
Function and homology information


deNEDDylase activity / protein deubiquitination / protein catabolic process / post-translational protein modification / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / ubiquitin binding / peptidase activity / UCH proteinases / Neddylation / ubiquitin-dependent protein catabolic process ...deNEDDylase activity / protein deubiquitination / protein catabolic process / post-translational protein modification / Synthesis of active ubiquitin: roles of E1 and E2 enzymes / ubiquitin binding / peptidase activity / UCH proteinases / Neddylation / ubiquitin-dependent protein catabolic process / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / protein ubiquitination / Golgi apparatus / nucleoplasm / cytosol / cytoplasm
Similarity search - Function
: / Ubiquitin carboxyl-terminal hydrolase family 1 cysteine active-site. / Peptidase C12, ubiquitin carboxyl-terminal hydrolase superfamily / Ubiquitin carboxyl-terminal hydrolase, family 1 / Ubiquitin carboxyl-terminal hydrolase (UCH) catalytic domain profile. / Peptidase C12, ubiquitin carboxyl-terminal hydrolase / Papain-like cysteine peptidase superfamily
Similarity search - Domain/homology
: / Ubiquitin carboxyl-terminal hydrolase isozyme L3
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 1.94 Å
AuthorsPatel, R. / Pannala, N. / Flaherty, D. / Das, C.
Funding support1items
OrganizationGrant numberCountry
Not funded
CitationJournal: Biorxiv / Year: 2026
Title: Identification, optimization, and structural elucidation of chloroacetamide scaffold as covalent inhibitors for Ubiquitin C-terminal Hydrolase L3
Authors: Beeralingappa, N.C. / Lu, M. / Patel, R. / Pannala, N. / Dhiman, A. / Heil, B.N. / Imhoff, R.D. / Smith, E.G. / Bahler, M.B. / Marsden, H.L. / Allen-Petersen, B.L. / Wendt, M.K. / Das, C. / Flaherty, D.P.
History
DepositionApr 30, 2026Deposition site: RCSB / Processing site: RCSB
Revision 1.0Aug 26, 2026Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Ubiquitin carboxyl-terminal hydrolase isozyme L3
B: Ubiquitin carboxyl-terminal hydrolase isozyme L3
C: Ubiquitin carboxyl-terminal hydrolase isozyme L3
D: Ubiquitin carboxyl-terminal hydrolase isozyme L3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)108,62018
Polymers106,5004
Non-polymers2,11914
Water5,495305
1
A: Ubiquitin carboxyl-terminal hydrolase isozyme L3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,2996
Polymers26,6251
Non-polymers6745
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
2
B: Ubiquitin carboxyl-terminal hydrolase isozyme L3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,1074
Polymers26,6251
Non-polymers4823
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
3
C: Ubiquitin carboxyl-terminal hydrolase isozyme L3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,0113
Polymers26,6251
Non-polymers3862
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
4
D: Ubiquitin carboxyl-terminal hydrolase isozyme L3
hetero molecules


Theoretical massNumber of molelcules
Total (without water)27,2035
Polymers26,6251
Non-polymers5784
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)48.029, 85.605, 124.078
Angle α, β, γ (deg.)90.00, 96.81, 90.00
Int Tables number4
Space group name H-MP1211

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Components

#1: Protein
Ubiquitin carboxyl-terminal hydrolase isozyme L3 / UCH-L3 / Ubiquitin thioesterase L3


Mass: 26625.031 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: UCHL3 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: P15374, ubiquitinyl hydrolase 1
#2: Chemical
ChemComp-A1DFD / 3-(2-chloroacetamido)-N-(pyridin-2-yl)benzamide


Mass: 289.717 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C14H12ClN3O2 / Feature type: SUBJECT OF INVESTIGATION
#3: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 10 / Source method: obtained synthetically / Formula: SO4
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 305 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY
Has protein modificationY

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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: 2.36 Å3/Da / Density % sol: 48 %
Crystal growTemperature: 298 K / Method: vapor diffusion, sitting drop / pH: 7 / Details: 200 mM Bis-Tris Propane 2.5 M Ammonium Sulfate

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: SSRL / Beamline: BL12-2 / Wavelength: 0.98 Å
DetectorType: DECTRIS EIGER2 XE 16M / Detector: PIXEL / Date: Dec 13, 2024
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.98 Å / Relative weight: 1
ReflectionResolution: 1.94→50 Å / Num. obs: 71476 / % possible obs: 98.2 % / Redundancy: 5.5 % / CC1/2: 0.995 / CC star: 0.999 / Rmerge(I) obs: 0.128 / Rpim(I) all: 0.058 / Rrim(I) all: 0.141 / Χ2: 1.158 / Net I/σ(I): 8.2
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique obsCC1/2CC starRpim(I) allRrim(I) allΧ2% possible all
1.95-2.025.31.21370250.5920.8620.561.341.19397
2.02-2.15.70.96772260.7470.9250.4231.0581.19499.7
2.1-2.25.70.6772330.8570.9610.2940.7331.19699.7
2.2-2.315.70.49871740.910.9760.2210.5461.18699.1
2.31-2.465.50.38371790.9350.9830.1720.4211.19699.2
2.46-2.655.40.28271820.9590.9890.1290.3111.15498.6
2.65-2.915.20.20968870.9720.9930.0980.2321.19894.9
2.91-3.335.80.13872480.9880.9970.0620.1521.12199.5
3.33-4.25.40.0971550.9930.9980.0420.11.04197.8
4.2-505.50.06771880.9960.9990.0310.0741.10296.5

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Processing

Software
NameVersionClassification
PHENIX(1.20.1_4487: ???)refinement
HKL-3000data scaling
HKL-3000data reduction
PHENIXphasing
PDB_EXTRACTdata extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 1.94→42.8 Å / SU ML: 0.25 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 28.71 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2437 3829 2.78 %
Rwork0.2036 --
obs0.2047 71476 95.59 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.1 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 1.94→42.8 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6744 0 126 305 7175
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0157052
X-RAY DIFFRACTIONf_angle_d2.0389585
X-RAY DIFFRACTIONf_dihedral_angle_d7.265922
X-RAY DIFFRACTIONf_chiral_restr0.1341056
X-RAY DIFFRACTIONf_plane_restr0.0141255
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
1.94-1.970.31081060.37183663X-RAY DIFFRACTION71
1.97-20.38841450.35554979X-RAY DIFFRACTION95
2-2.020.41211320.32965098X-RAY DIFFRACTION99
2.02-2.050.35761430.31685120X-RAY DIFFRACTION99
2.05-2.080.36631500.31565153X-RAY DIFFRACTION99
2.08-2.110.32021430.29485070X-RAY DIFFRACTION99
2.11-2.150.28711580.27825147X-RAY DIFFRACTION99
2.15-2.190.29981280.26255191X-RAY DIFFRACTION98
2.19-2.230.34921480.24885094X-RAY DIFFRACTION99
2.23-2.270.311700.24175057X-RAY DIFFRACTION98
2.27-2.320.25861440.22625116X-RAY DIFFRACTION98
2.32-2.370.27491300.22665091X-RAY DIFFRACTION98
2.37-2.420.23721480.22715074X-RAY DIFFRACTION98
2.42-2.480.23481540.2265081X-RAY DIFFRACTION98
2.48-2.550.27631470.22385087X-RAY DIFFRACTION97
2.55-2.620.23131350.21794973X-RAY DIFFRACTION97
2.62-2.710.24711460.22654922X-RAY DIFFRACTION94
2.71-2.80.29841470.23964630X-RAY DIFFRACTION89
2.8-2.920.25431370.21794942X-RAY DIFFRACTION96
2.92-3.050.23051410.2135147X-RAY DIFFRACTION98
3.05-3.210.26661470.20225079X-RAY DIFFRACTION97
3.21-3.410.24581480.19495020X-RAY DIFFRACTION97
3.41-3.670.30261410.17224874X-RAY DIFFRACTION94
3.67-4.040.18251270.164863X-RAY DIFFRACTION93
4.04-4.630.16571470.154719X-RAY DIFFRACTION92
4.63-5.830.17421220.16524757X-RAY DIFFRACTION91
5.83-42.80.24131450.18385131X-RAY DIFFRACTION99
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)
10.5566-0.3705-1.17835.3908-0.35344.7261-0.14180.0601-0.09970.09880.20670.45950.252-0.5303-0.03710.2638-0.10740.04240.4487-0.07320.3705-20.3867-0.9288-63.7544
21.3778-0.2139-0.36422.84860.38341.26130.0074-0.0359-0.01750.04470.0568-0.1161-0.0020.0388-0.05220.1742-0.01730.01970.32090.00070.2805-12.21688.5366-50.9696
30.86920.2037-1.27495.08170.22523.98780.08780.24830.2387-0.46260.03160.1694-0.3972-0.2353-0.15140.2337-0.0066-0.02520.40120.04580.2788-18.725614.9477-65.8334
43.43820.0739-1.10942.95630.18792.8269-0.0674-0.002-0.20320.14980.02610.05570.24220.00780.06210.20980.00590.03660.217400.2541-11.72514.1369-52.2627
52.880.58240.90139.1314-1.18630.5252-0.3299-0.33390.4395-0.1327-0.1694-0.65890.1747-0.33540.68660.5432-0.0046-0.14380.343-0.01980.5187-26.258723.898-20.1738
62.3872-1.81310.84343.8728-0.43532.3382-0.3321-0.34140.27760.80250.07110.1305-0.4192-0.11580.28260.5688-0.01480.05770.2682-0.0630.3807-37.735727.2332-22.0406
73.6085-3.17193.6573.8978-2.65338.63250.06370.1631-0.29560.1957-0.02260.31760.4477-0.1352-0.10620.2859-0.03690.09310.2149-0.05690.4042-39.52857.7105-39.4717
81.7351-1.10740.22363.014-0.01332.3587-0.03140.14040.7707-0.3068-0.1178-0.8946-0.41980.28850.15660.3584-0.06210.05320.27260.0670.5919-28.494928.7491-37.5144
92.2253-0.4362-0.78643.5049-0.3693.6260.063-0.03390.5794-0.0332-0.1488-1.0312-0.0720.62260.08860.3056-0.04180.0530.31670.00060.5734-24.664520.8438-32.857
105.0811-2.1812-0.96334.53550.13183.8492-0.06770.49260.03360.0397-0.13940.153-0.0111-0.2277-0.010.2609-0.01170.04210.2178-0.00730.3082-40.49421.557-38.3518
114.3018-3.58421.40673.2924-2.51685.8712-0.4923-0.4744-0.23360.11420.51850.3382-0.1489-0.44940.01320.43740.0030.15430.3674-0.02140.4344-45.623721.0167-27.7841
123.8409-1.23590.68772.5853-1.75815.05-0.30640.0311-0.02910.71050.2076-0.2597-0.1-0.07130.11940.45160.04990.17570.23620.00860.3308-37.500817.4722-21.8987
139.80020.5742-5.38821.5552-0.43033.06190.67050.19920.50970.2361-0.05890.4124-0.9011-0.3946-0.24410.63020.06260.22420.35880.09770.5443-53.38320.047-6.4712
142.2865-0.80420.94012.98881.38453.8563-0.6648-0.3629-0.02310.54950.29870.0906-0.5556-0.0144-0.57790.96360.14350.56110.38310.00740.5707-47.22838.00683.9208
150.6740.11330.10192.5644-1.95153.12160.1329-0.0090.13540.0755-0.0689-0.37750.06190.3985-0.03380.5776-0.04760.11270.3011-0.0490.4363-30.9339-6.6743-6.1324
161.42070.25320.66192.6390.31884.2045-0.0572-0.287-0.19480.47640.0320.57260.4862-0.20090.02120.71640.00010.23880.3420.06140.4722-48.2657-13.69871.2576
173.22890.28271.3933.28141.47534.725-0.0276-0.20610.05810.5487-0.0660.01120.13780.00130.09660.6812-0.01710.15060.28490.04110.3681-37.7162-5.58550.0881
182.9093-0.0401-0.13054.6384-0.94263.63820.1538-0.23430.23080.304-0.0131-0.1764-0.0730.1041-0.18540.6844-0.02070.17070.2366-0.0050.4036-36.48374.2614-3.3755
194.2029-0.65292.52452.9349-3.30228.18940.23680.47190.4596-0.1123-0.581-1.2761.04330.80630.27990.60870.12840.0390.43660.01470.6311-16.6737-21.782-31.0432
201.02260.81821.6145.26830.06382.8965-0.09670.1201-0.38580.5569-0.0156-1.30921.46530.5092-0.15421.11720.11360.17060.3552-0.06340.7258-21.465-31.5093-31.9743
212.1098-1.4805-0.36464.22091.6561.61830.02880.13830.00820.1025-0.14280.48120.1522-0.19670.09030.4509-0.07030.14940.28470.00410.4012-34.2941-16.0519-36.8937
222.981.20070.26333.07870.3533.4444-0.0947-0.2573-0.48520.82830.08270.24920.9519-0.0360.01881.0696-0.05430.2270.31630.01920.5431-31.1087-26.7651-21.2204
231.3243-0.8339-0.45553.46560.45554.2161-0.1020.0121-0.18390.3442-0.03340.09550.453-0.10920.07560.5395-0.06490.23430.2922-0.03290.4911-31.546-19.172-30.9987
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 4 through 38 )
2X-RAY DIFFRACTION2chain 'A' and (resid 39 through 94 )
3X-RAY DIFFRACTION3chain 'A' and (resid 95 through 161 )
4X-RAY DIFFRACTION4chain 'A' and (resid 162 through 230 )
5X-RAY DIFFRACTION5chain 'B' and (resid 5 through 22 )
6X-RAY DIFFRACTION6chain 'B' and (resid 23 through 59 )
7X-RAY DIFFRACTION7chain 'B' and (resid 60 through 76 )
8X-RAY DIFFRACTION8chain 'B' and (resid 77 through 130 )
9X-RAY DIFFRACTION9chain 'B' and (resid 131 through 176 )
10X-RAY DIFFRACTION10chain 'B' and (resid 177 through 197 )
11X-RAY DIFFRACTION11chain 'B' and (resid 198 through 215 )
12X-RAY DIFFRACTION12chain 'B' and (resid 216 through 230 )
13X-RAY DIFFRACTION13chain 'C' and (resid 5 through 23 )
14X-RAY DIFFRACTION14chain 'C' and (resid 24 through 48 )
15X-RAY DIFFRACTION15chain 'C' and (resid 49 through 76 )
16X-RAY DIFFRACTION16chain 'C' and (resid 77 through 161 )
17X-RAY DIFFRACTION17chain 'C' and (resid 162 through 200 )
18X-RAY DIFFRACTION18chain 'C' and (resid 201 through 230 )
19X-RAY DIFFRACTION19chain 'D' and (resid 5 through 22 )
20X-RAY DIFFRACTION20chain 'D' and (resid 23 through 34 )
21X-RAY DIFFRACTION21chain 'D' and (resid 35 through 76 )
22X-RAY DIFFRACTION22chain 'D' and (resid 77 through 130 )
23X-RAY DIFFRACTION23chain 'D' and (resid 131 through 230 )

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