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- PDB-3ecq: Endo-alpha-N-acetylgalactosaminidase from Streptococcus pneumonia... -

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Basic information

Entry
Database: PDB / ID: 3ecq
TitleEndo-alpha-N-acetylgalactosaminidase from Streptococcus pneumoniae: SeMet structure
ComponentsEndo-alpha-N-acetylgalactosaminidase
KeywordsHYDROLASE / Distorted (Beta/Alpha)8 (TIM) barrel glycoside hydrolase domain / Cell wall / Peptidoglycan-anchor / Secreted
Function / homology
Function and homology information


endo-alpha-N-acetylgalactosaminidase / : / endo-alpha-N-acetylgalactosaminidase activity / cell wall / : / metabolic process / carbohydrate binding / extracellular region / membrane / metal ion binding
Similarity search - Function
Jelly Rolls - #870 / Helix Hairpins - #660 / Glycosyl hydrolase 101, beta-sandwich domain / Galactose-binding domain-like / Endo-alpha-N-acetylgalactosaminidase, N-terminal / Galactose mutarotase-like fold domain / Endo-alpha-N-acetylgalactosaminidase / Glycosyl hydrolase 101 beta sandwich domain / Galactose-binding domain-like / Endo-alpha-N-acetylgalactosaminidase N-terminal domain ...Jelly Rolls - #870 / Helix Hairpins - #660 / Glycosyl hydrolase 101, beta-sandwich domain / Galactose-binding domain-like / Endo-alpha-N-acetylgalactosaminidase, N-terminal / Galactose mutarotase-like fold domain / Endo-alpha-N-acetylgalactosaminidase / Glycosyl hydrolase 101 beta sandwich domain / Galactose-binding domain-like / Endo-alpha-N-acetylgalactosaminidase N-terminal domain / Galactose mutarotase-like fold domain / Endo-alpha-N-acetylgalactosaminidase / Beta-galactosidase; Chain A, domain 5 - #10 / Glycoside hydrolase-type carbohydrate-binding / YSIRK type signal peptide / F5/8 type C domain / Coagulation factor 5/8 C-terminal domain / YSIRK Gram-positive signal peptide / Beta-galactosidase; Chain A, domain 5 / Helix Hairpins / Gram-positive cocci surface proteins LPxTG motif profile. / LPXTG cell wall anchor domain / Galactose-binding domain-like / Golgi alpha-mannosidase II / Glycosyl hydrolase, all-beta / Helix non-globular / Galactose-binding-like domain superfamily / Distorted Sandwich / Special / Glycosidases / Jelly Rolls / TIM Barrel / Alpha-Beta Barrel / Immunoglobulin-like / Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
Endo-alpha-N-acetylgalactosaminidase
Similarity search - Component
Biological speciesStreptococcus pneumoniae (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / SAD / Resolution: 2.9 Å
AuthorsCaines, M.E.C. / Zhu, H. / Vuckovic, M. / Strynadka, N.C.J.
CitationJournal: J.Biol.Chem. / Year: 2008
Title: The structural basis for T-antigen hydrolysis by Streptococcus pneumoniae: a target for structure-based vaccine design.
Authors: Caines, M.E. / Zhu, H. / Vuckovic, M. / Willis, L.M. / Withers, S.G. / Wakarchuk, W.W. / Strynadka, N.C.
History
DepositionSep 1, 2008Deposition site: RCSB / Processing site: RCSB
Revision 1.0Sep 9, 2008Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Non-polymer description / Version format compliance

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Endo-alpha-N-acetylgalactosaminidase
B: Endo-alpha-N-acetylgalactosaminidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)343,60910
Polymers343,2192
Non-polymers3908
Water1,74797
1
A: Endo-alpha-N-acetylgalactosaminidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)171,8055
Polymers171,6091
Non-polymers1954
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: Endo-alpha-N-acetylgalactosaminidase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)171,8055
Polymers171,6091
Non-polymers1954
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)210.611, 158.209, 112.436
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number18
Space group name H-MP21212
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22B
13A
23B
14A
24B
15A
25B
16A
26B
17A
27B

NCS domain segments:

Refine code: 2

Dom-IDComponent-IDEns-IDBeg label comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111THRGLUAA122 - 30786 - 271
211THRGLUBB122 - 30786 - 271
112THRSERAA330 - 354294 - 318
212THRSERBB330 - 354294 - 318
122LEUGLNAA360 - 601324 - 565
222LEUGLNBB360 - 601324 - 565
113GLYGLUAA602 - 655566 - 619
213GLYGLUBB602 - 655566 - 619
123HISSERAA661 - 721625 - 685
223HISSERBB661 - 721625 - 685
133GLYGLUAA730 - 796694 - 760
233GLYGLUBB730 - 796694 - 760
143GLYHISAA800 - 808764 - 772
243GLYHISBB800 - 808764 - 772
153GLYTYRAA818 - 859782 - 823
253GLYTYRBB818 - 859782 - 823
163ASNPHEAA874 - 893838 - 857
263ASNPHEBB874 - 893838 - 857
114THRTHRAA894 - 907858 - 871
214THRTHRBB894 - 907858 - 871
124LYSARGAA914 - 1043878 - 1007
224LYSARGBB914 - 1043878 - 1007
115HISGLUAA1058 - 12131022 - 1177
215HISGLUBB1058 - 12131022 - 1177
116ASNSERAA1214 - 13391178 - 1303
216ASNSERBB1214 - 13391178 - 1303
126LEUGLUAA1349 - 14171313 - 1381
226LEUGLUBB1349 - 14171313 - 1381
117THRLEUAA1421 - 14341385 - 1398
217THRLEUBB1421 - 14341385 - 1398
127LYSILEAA1451 - 14761415 - 1440
227LYSILEBB1451 - 14761415 - 1440

NCS ensembles :
ID
1
2
3
4
5
6
7

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Components

#1: Protein Endo-alpha-N-acetylgalactosaminidase / protein spr0328


Mass: 171609.422 Da / Num. of mol.: 2 / Fragment: UNP residues 40-1567
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Streptococcus pneumoniae (bacteria) / Strain: R6 / Gene: spr0328 / Plasmid: pCWori+ / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: Q8DR60, endo-alpha-N-acetylgalactosaminidase
#2: Chemical
ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: Ca
#3: Chemical ChemComp-NA / SODIUM ION


Mass: 22.990 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Na
#4: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C3H8O3
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 97 / 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: 2.73 Å3/Da / Density % sol: 54.93 %
Crystal growTemperature: 294 K / Method: vapor diffusion, sitting drop
Details: 29% (w/v) PEG MME 2000, 0.2 M lithium citrate, 0.1 M ammonium acetate, VAPOR DIFFUSION, SITTING DROP, temperature 294K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 8.2.2 / Wavelength: 0.9795 Å
DetectorType: ADSC QUANTUM 315 / Detector: CCD / Date: Sep 30, 2006
RadiationMonochromator: Double crystal, Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 2.9→112.509 Å / Num. obs: 83445 / % possible obs: 99.4 % / Redundancy: 6.8 % / Biso Wilson estimate: 71.87 Å2 / Rmerge(I) obs: 0.11 / Rsym value: 0.11 / Net I/σ(I): 6.13
Reflection shellResolution: 2.9→3.06 Å / Redundancy: 6.3 % / Rmerge(I) obs: 0.592 / Mean I/σ(I) obs: 2.4 / Num. measured all: 74051 / Num. unique all: 11781 / Rsym value: 0.592 / % possible all: 97.4

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Phasing

PhasingMethod: SAD

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Processing

Software
NameVersionClassificationNB
MOSFLMdata reduction
SCALA3.2.17data scaling
SHARPphasing
SOLOMONphasing
REFMACrefinement
PDB_EXTRACT3.006data extraction
RefinementMethod to determine structure: SAD / Resolution: 2.9→112.509 Å / Cor.coef. Fo:Fc: 0.941 / Cor.coef. Fo:Fc free: 0.907 / WRfactor Rfree: 0.225 / WRfactor Rwork: 0.181 / Occupancy max: 1 / Occupancy min: 0.5 / FOM work R set: 0.863 / SU B: 30.846 / SU ML: 0.268 / SU R Cruickshank DPI: 0.313 / SU Rfree: 0.38 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.36
Stereochemistry target values: MAXIMUM LIKELIHOOD WITH PHASES
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. THE ATOM RECORDS CONTAIN FULL B-FACTORS. THE ANISOU RECORDS RESULT FROM TLS REFINEMENT
RfactorNum. reflection% reflectionSelection details
Rfree0.235 4164 5 %RANDOM
Rwork0.19 ---
obs0.192 83381 99.37 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å / Solvent model: MASK
Displacement parametersBiso max: 159.15 Å2 / Biso mean: 57.319 Å2 / Biso min: 13.08 Å2
Baniso -1Baniso -2Baniso -3
1--0.99 Å20 Å20 Å2
2--0.91 Å20 Å2
3---0.08 Å2
Refinement stepCycle: LAST / Resolution: 2.9→112.509 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms21250 0 18 97 21365
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.02221735
X-RAY DIFFRACTIONr_bond_other_d0.0010.0214463
X-RAY DIFFRACTIONr_angle_refined_deg1.2251.92629429
X-RAY DIFFRACTIONr_angle_other_deg0.823335240
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.58752679
X-RAY DIFFRACTIONr_dihedral_angle_2_deg38.5525.0871091
X-RAY DIFFRACTIONr_dihedral_angle_3_deg16.777153681
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.6451594
X-RAY DIFFRACTIONr_chiral_restr0.0710.23139
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.0224571
X-RAY DIFFRACTIONr_gen_planes_other0.0010.024427
X-RAY DIFFRACTIONr_nbd_refined0.2070.24296
X-RAY DIFFRACTIONr_nbd_other0.1890.214667
X-RAY DIFFRACTIONr_nbtor_refined0.1860.210506
X-RAY DIFFRACTIONr_nbtor_other0.0860.212015
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1360.2555
X-RAY DIFFRACTIONr_xyhbond_nbd_other0.0220.21
X-RAY DIFFRACTIONr_metal_ion_refined0.1870.218
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.1250.215
X-RAY DIFFRACTIONr_symmetry_vdw_other0.1680.245
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.3310.23
X-RAY DIFFRACTIONr_mcbond_it0.2861.513241
X-RAY DIFFRACTIONr_mcbond_other0.081.55536
X-RAY DIFFRACTIONr_mcangle_it0.625221276
X-RAY DIFFRACTIONr_scbond_it1.31338901
X-RAY DIFFRACTIONr_scangle_it2.0694.58152
Refine LS restraints NCS

Dom-ID: 1 / Auth asym-ID: A / Refine-ID: X-RAY DIFFRACTION

Ens-IDNumberTypeRms dev position (Å)Weight position
11080TIGHT POSITIONAL0.030.05
11366MEDIUM POSITIONAL0.410.5
11080TIGHT THERMAL0.030.5
11366MEDIUM THERMAL0.22
21578TIGHT POSITIONAL0.030.05
21938MEDIUM POSITIONAL0.340.5
21578TIGHT THERMAL0.040.5
21938MEDIUM THERMAL0.282
31481TIGHT POSITIONAL0.040.05
31927MEDIUM POSITIONAL0.330.5
31481TIGHT THERMAL0.050.5
31927MEDIUM THERMAL0.322
4851TIGHT POSITIONAL0.030.05
41119MEDIUM POSITIONAL0.320.5
4851TIGHT THERMAL0.040.5
41119MEDIUM THERMAL0.262
5923TIGHT POSITIONAL0.030.05
51098MEDIUM POSITIONAL0.360.5
5923TIGHT THERMAL0.050.5
51098MEDIUM THERMAL0.312
61143TIGHT POSITIONAL0.030.05
61456MEDIUM POSITIONAL0.280.5
61143TIGHT THERMAL0.050.5
61456MEDIUM THERMAL0.312
7239TIGHT POSITIONAL0.030.05
7277MEDIUM POSITIONAL0.430.5
7239TIGHT THERMAL0.030.5
7277MEDIUM THERMAL0.22
LS refinement shellResolution: 2.9→2.975 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.403 298 -
Rwork0.351 5616 -
all-5914 -
obs-5616 96.46 %
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)
13.2285-0.115-1.16686.7896-1.10694.41390.13010.23910.09670.11490.03260.7406-0.543-0.3907-0.16280.07170.15830.1009-0.0694-0.0179-0.080954.73362.306313.117
21.6930.3101-0.38321.22980.25781.39060.0709-0.18370.12880.04820.1041-0.5938-0.16560.3898-0.175-0.2172-0.0037-0.027-0.1376-0.26350.232985.959341.469933.2672
30.33380.1881-0.36170.93370.61131.19460.0620.1630.0279-0.15560.0645-0.0217-0.0348-0.3328-0.1265-0.15980.04550.0558-0.2092-0.0939-0.105458.128528.221619.3914
41.43120.10470.4261.85351.25851.66460.06810.2473-0.0721-0.35090.131-0.5428-0.07050.2723-0.1991-0.09040.03380.2284-0.1148-0.1632-0.038273.632721.9381-3.4228
51.9329-0.6788-0.72561.79050.01041.88110.02160.2941-0.1049-0.22570.0716-0.18760.2866-0.0826-0.0932-0.1689-0.10570.09-0.2081-0.0948-0.360534.864610.81313.8767
61.53210.36660.14551.85260.76392.0770.0139-0.02520.21520.1069-0.01930.2545-0.0137-0.44220.0055-0.2839-0.0110.1307-0.1326-0.0214-0.285423.963529.541520.2595
712.20321.7121.16149.79610.293114.0906-0.50380.13761.1371-0.47930.01270.3492-1.0693-0.96050.49110.01690.09310.12670.37380.07680.369-1.081432.064520.4993
82.60811.18270.34463.43330.90313.20380.01250.2775-0.61350.50050.115-0.68570.27350.4725-0.12750.37120.1132-0.05870.3353-0.12920.5649.406823.1371-43.5874
92.28360.2269-0.11711.9585-1.27132.0708-0.05110.05740.0066-0.17640.08350.36150.3337-0.4042-0.0325-0.1684-0.03390.0033-0.1637-0.0031-0.155721.180945.1892-22.9321
100.86990.66340.1052.2155-1.33241.60340.04360.37980.0178-0.2786-0.0972-0.49990.04920.46110.0536-0.12110.0548-0.0055-0.08160.0608-0.129549.709858.0662-36.2605
112.59191.271-0.86692.1848-1.36522.1782-0.21960.81950.5824-0.64760.4060.45720.3461-0.5118-0.18640.1077-0.0343-0.15910.26220.2551-0.111434.487265.6795-58.7807
121.691-0.90350.39021.60150.42541.65090.1770.3904-0.0932-0.250.19390.1606-0.37910.0594-0.3709-0.1119-0.04550.0817-0.1649-0.0149-0.260473.79574.9191-51.1443
132.27540.5348-0.28411.4421-0.61292.39260.30070.0389-0.69230.06050.0765-0.28890.13580.2629-0.3772-0.2831-0.0281-0.1734-0.174-0.10510.070683.664455.0316-35.5462
146.7846-0.601-0.85737.7689-2.24255.44190.07060.561-1.0351-0.6792-0.17370.46530.77890.25580.10310.05350.1289-0.22030.547-0.3460.3985106.546345.9045-38.6047
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
1X-RAY DIFFRACTION1AA119 - 30783 - 271
2X-RAY DIFFRACTION2AA319 - 601283 - 565
3X-RAY DIFFRACTION3AA602 - 893566 - 857
4X-RAY DIFFRACTION4AA894 - 1057858 - 1021
5X-RAY DIFFRACTION5AA1058 - 12131022 - 1177
6X-RAY DIFFRACTION6AA1214 - 14171178 - 1381
7X-RAY DIFFRACTION7AA1418 - 14761382 - 1440
8X-RAY DIFFRACTION8BB122 - 30786 - 271
9X-RAY DIFFRACTION9BB318 - 601282 - 565
10X-RAY DIFFRACTION10BB602 - 893566 - 857
11X-RAY DIFFRACTION11BB894 - 1057858 - 1021
12X-RAY DIFFRACTION12BB1058 - 12131022 - 1177
13X-RAY DIFFRACTION13BB1214 - 14171178 - 1381
14X-RAY DIFFRACTION14BB1418 - 14811382 - 1445

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