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- PDB-5hup: Crystal Structure of NadC from Streptococcus pyogenes -

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Basic information

Entry
Database: PDB / ID: 5hup
TitleCrystal Structure of NadC from Streptococcus pyogenes
ComponentsNicotinate-nucleotide pyrophosphorylase (Carboxylating)
KeywordsTRANSFERASE / Quinolinate phosphoribosyltransferase
Function / homology
Function and homology information


nicotinate-nucleotide diphosphorylase (carboxylating) / nicotinate-nucleotide diphosphorylase (carboxylating) activity / NAD biosynthetic process
Similarity search - Function
Nicotinate-nucleotide pyrophosphorylase / Nicotinate-nucleotide pyrophosphorylase/Putative pyrophosphorylase ModD / Quinolinate phosphoribosyl transferase, N-terminal / Quinolinate phosphoribosyl transferase, N-terminal domain / Quinolinate phosphoribosyl transferase, N-terminal domain / Quinolinate phosphoribosyl transferase, C-terminal / Quinolinate phosphoribosyl transferase, N-terminal domain superfamily / Quinolinate phosphoribosyl transferase, C-terminal domain / Nicotinate phosphoribosyltransferase-like, C-terminal / Aldehyde Oxidoreductase; domain 3 ...Nicotinate-nucleotide pyrophosphorylase / Nicotinate-nucleotide pyrophosphorylase/Putative pyrophosphorylase ModD / Quinolinate phosphoribosyl transferase, N-terminal / Quinolinate phosphoribosyl transferase, N-terminal domain / Quinolinate phosphoribosyl transferase, N-terminal domain / Quinolinate phosphoribosyl transferase, C-terminal / Quinolinate phosphoribosyl transferase, N-terminal domain superfamily / Quinolinate phosphoribosyl transferase, C-terminal domain / Nicotinate phosphoribosyltransferase-like, C-terminal / Aldehyde Oxidoreductase; domain 3 / Aldolase class I / Aldolase-type TIM barrel / TIM Barrel / Alpha-Beta Barrel / Alpha-Beta Complex / Alpha Beta
Similarity search - Domain/homology
Quinolinate phosphoribosyltransferase [decarboxylating]
Similarity search - Component
Biological speciesStreptococcus pyogenes serotype M4 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / FOURIER SYNTHESIS / Resolution: 3.42 Å
AuthorsBooth, W.T. / Chruszcz, M.
CitationJournal: FEBS J. / Year: 2017
Title: Streptococcus pyogenes quinolinate-salvage pathway-structural and functional studies of quinolinate phosphoribosyl transferase and NH3 -dependent NAD(+) synthetase.
Authors: Booth, W.T. / Morris, T.L. / Mysona, D.P. / Shah, M.J. / Taylor, L.K. / Karlin, T.W. / Clary, K. / Majorek, K.A. / Offermann, L.R. / Chruszcz, M.
History
DepositionJan 27, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Jan 25, 2017Provider: repository / Type: Initial release
Revision 1.1Jul 12, 2017Group: Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year
Revision 1.2Aug 23, 2017Group: Database references / Category: citation
Item: _citation.journal_volume / _citation.page_first ..._citation.journal_volume / _citation.page_first / _citation.page_last / _citation.title
Revision 1.3Sep 27, 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 / struct_ncs_dom_lim
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
B: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
C: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
E: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
F: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
H: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)208,61523
Polymers206,9826
Non-polymers1,63317
Water00
1
A: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
C: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
E: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
hetero molecules

A: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
C: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
E: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)208,71124
Polymers206,9826
Non-polymers1,72918
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation4_555x,-y,-z1
Buried area28430 Å2
ΔGint-367 kcal/mol
Surface area58330 Å2
MethodPISA
2
B: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
F: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
H: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
hetero molecules

B: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
F: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
H: Nicotinate-nucleotide pyrophosphorylase (Carboxylating)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)208,51922
Polymers206,9826
Non-polymers1,53716
Water0
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation3_454-x-1,y,-z-1/21
Buried area29630 Å2
ΔGint-337 kcal/mol
Surface area57480 Å2
MethodPISA
Unit cell
Length a, b, c (Å)108.149, 188.816, 222.181
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number20
Space group name H-MC2221
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
12A
22C
13A
23E
14A
24F
15A
25H
16B
26C
17B
27E
18B
28F
19B
29H
110C
210E
111C
211F
112C
212H
113E
213F
114E
214H
115F
215H

NCS domain segments:

Component-ID: _ / Refine code: _

Dom-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11SERSERASPASPAA5 - 28930 - 314
21SERSERASPASPBB5 - 28930 - 314
12LEULEULEULEUAA8 - 28833 - 313
22LEULEULEULEUCC8 - 28833 - 313
13SERSERTYRTYRAA5 - 28730 - 312
23SERSERTYRTYRED5 - 28730 - 312
14THRTHRLEULEUAA6 - 28831 - 313
24THRTHRLEULEUFE6 - 28831 - 313
15SERSERASPASPAA5 - 28930 - 314
25SERSERASPASPHF5 - 28930 - 314
16LEULEULEULEUBB8 - 28833 - 313
26LEULEULEULEUCC8 - 28833 - 313
17SERSERTYRTYRBB5 - 28730 - 312
27SERSERTYRTYRED5 - 28730 - 312
18THRTHRLEULEUBB6 - 28831 - 313
28THRTHRLEULEUFE6 - 28831 - 313
19SERSERASPASPBB5 - 28930 - 314
29SERSERASPASPHF5 - 28930 - 314
110LEULEUTYRTYRCC8 - 28733 - 312
210LEULEUTYRTYRED8 - 28733 - 312
111LEULEUASPASPCC8 - 28933 - 314
211LEULEUASPASPFE8 - 28933 - 314
112LEULEULEULEUCC8 - 28833 - 313
212LEULEULEULEUHF8 - 28833 - 313
113THRTHRTYRTYRED6 - 28731 - 312
213THRTHRTYRTYRFE6 - 28731 - 312
114SERSERTYRTYRED5 - 28730 - 312
214SERSERTYRTYRHF5 - 28730 - 312
115THRTHRLEULEUFE6 - 28831 - 313
215THRTHRLEULEUHF6 - 28831 - 313

NCS ensembles :
ID
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15

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Components

#1: Protein
Nicotinate-nucleotide pyrophosphorylase (Carboxylating)


Mass: 34496.980 Da / Num. of mol.: 6 / Fragment: UNP residues 10-299
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Streptococcus pyogenes serotype M4 (strain MGAS10750) (bacteria)
Strain: MGAS10750 / Gene: nadC, MGAS10750_Spy1186 / Plasmid: pjExpress411 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: Q1J647, nicotinate-nucleotide diphosphorylase (carboxylating)
#2: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 17 / Source method: obtained synthetically / Formula: SO4

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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: 2.74 Å3/Da / Density % sol: 55.11 %
Crystal growTemperature: 293 K / Method: vapor diffusion / pH: 6.5
Details: 0.48 M Ammonium Sulfate, 25% PEG 6000, 0.1 M BIS-TRIS

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 19-BM / Wavelength: 0.97919 Å
DetectorType: ADSC QUANTUM 210r / Detector: CCD / Date: Oct 25, 2015
RadiationMonochromator: Si 111 / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97919 Å / Relative weight: 1
ReflectionResolution: 3.4→20 Å / Num. obs: 25907 / % possible obs: 81.6 % / Observed criterion σ(I): -3 / Redundancy: 3.5 % / Rmerge(I) obs: 0.107 / Rsym value: 0.107 / Net I/σ(I): 15.4
Reflection shellResolution: 3.4→3.46 Å / Redundancy: 3.6 % / Rmerge(I) obs: 0.606 / Mean I/σ(I) obs: 3.3 / % possible all: 95.2

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Processing

Software
NameVersionClassification
REFMAC5.8.0135refinement
HKL-2000data scaling
HKL-2000data reduction
REFMAC5.8.0135phasing
RefinementMethod to determine structure: FOURIER SYNTHESIS
Starting model: 5HUO
Resolution: 3.42→20 Å / Cor.coef. Fo:Fc: 0.884 / Cor.coef. Fo:Fc free: 0.817 / SU B: 81.17 / SU ML: 0.53 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.817 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.29354 1221 5 %RANDOM
Rwork0.23279 ---
obs0.23569 23334 78.86 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 86.544 Å2
Baniso -1Baniso -2Baniso -3
1-5.39 Å20 Å20 Å2
2---1.95 Å20 Å2
3----3.43 Å2
Refinement stepCycle: 1 / Resolution: 3.42→20 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms12666 0 85 0 12751
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0130.01912923
X-RAY DIFFRACTIONr_bond_other_d0.010.0212190
X-RAY DIFFRACTIONr_angle_refined_deg1.5581.95817540
X-RAY DIFFRACTIONr_angle_other_deg1.814327787
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.97851696
X-RAY DIFFRACTIONr_dihedral_angle_2_deg32.74123.659533
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.831152042
X-RAY DIFFRACTIONr_dihedral_angle_4_deg17.9531583
X-RAY DIFFRACTIONr_chiral_restr0.080.22116
X-RAY DIFFRACTIONr_gen_planes_refined0.0090.0214823
X-RAY DIFFRACTIONr_gen_planes_other0.0080.022976
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_it2.8445.4526805
X-RAY DIFFRACTIONr_mcbond_other2.8445.4526804
X-RAY DIFFRACTIONr_mcangle_it4.7968.1788494
X-RAY DIFFRACTIONr_mcangle_other4.7968.1788495
X-RAY DIFFRACTIONr_scbond_it3.2615.756118
X-RAY DIFFRACTIONr_scbond_other3.2015.6686048
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other5.4798.3888944
X-RAY DIFFRACTIONr_long_range_B_refined10.77351.38151284
X-RAY DIFFRACTIONr_long_range_B_other10.77351.38151285
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION / Type: interatomic distance / Weight position: 0.05

Ens-IDDom-IDAuth asym-IDNumberRms dev position (Å)
11A320780.08
12B320780.08
21A273060.1
22C273060.1
31A308600.09
32E308600.09
41A330360.06
42F330360.06
51A318120.09
52H318120.09
61B272120.1
62C272120.1
71B310920.09
72E310920.09
81B324980.07
82F324980.07
91B318620.09
92H318620.09
101C260600.12
102E260600.12
111C274420.1
112F274420.1
121C275180.09
122H275180.09
131E311400.09
132F311400.09
141E303760.11
142H303760.11
151F319180.08
152H319180.08
LS refinement shellResolution: 3.42→3.506 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.37 116 -
Rwork0.298 1767 -
obs--85.51 %
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)
10.7120.1680.34924.3984-0.57491.0984-0.04830.06880.0044-0.42750.1038-0.40650.26320.2497-0.05550.12720.05550.02590.3049-0.06880.0762-45.502-8.646-12.773
21.20360.8491-0.15654.54821.45371.76480.0687-0.17520.29190.1483-0.03640.1236-0.0528-0.0263-0.03230.0896-0.09740.02510.2599-0.04010.0814-51.71418.519-6.753
32.96074.6453-1.24819.2419-3.99952.7017-0.061-0.07790.0132-0.0841-0.022-0.02220.03280.02320.08290.15640.11260.10750.27830.01340.3149-43.223-42.697-47.19
40.24320.4381-0.47643.2517-1.94323.07010.021-0.1562-0.0728-0.0156-0.106-0.46730.11130.39110.0850.013-0.0071-0.01180.23910.02280.2871-27.232-32.702-64.085
54.55251.06110.5131.0557-0.16972.9170.0808-0.1917-0.3516-0.2528-0.2562-0.28890.1480.57990.17540.1362-0.04610.08040.50970.03540.4609-14.8129.022-19.902
60.23820.2433-0.46520.6185-0.15961.25940.0986-0.4524-0.2946-0.0361-0.4483-0.3766-0.36811.08510.34970.1529-0.3762-0.08951.48650.35720.76987.56413.602-3.844
73.6423-0.5071.11311.8067-1.08921.39850.178-0.5305-0.20390.4807-0.3748-0.4344-0.56790.74780.19680.4135-0.5426-0.17161.04470.07020.4216-12.20422.2935.599
82.45531.20361.9840.95980.32594.14970.2738-0.1174-0.01970.1524-0.0813-0.2558-0.22820.3144-0.19250.1808-0.2050.14290.3561-0.07470.4421-28.33327.387-18.224
98.1779-1.2052-1.21010.65160.53352.9009-0.0570.1576-0.5523-0.21280.01060.2383-0.03160.00160.04630.1426-0.0049-0.01880.0582-0.03960.2824-66.623-18.431-62.525
105.1752-0.13760.46790.9111-0.02240.36880.05090.11310.089-0.0439-0.0151-0.1075-0.2399-0.0381-0.03580.1976-0.00750.050.2009-0.02240.0447-45.907-3.625-65.414
110.7888-0.8919-0.11633.3978-0.82181.80070.05190.06320.0157-0.36510.02270.04460.10520.1114-0.07450.05480.0153-0.01110.08870.04150.2092-45.45-36.964-77.518
120.808-0.2208-0.20981.7007-1.51772.3021-0.05720.0014-0.0173-0.01690.00990.08360.17720.29420.04730.08310.05630.02520.1506-0.02310.2335-41.132-60.626-60.006
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A5 - 137
2X-RAY DIFFRACTION2A138 - 289
3X-RAY DIFFRACTION3B5 - 39
4X-RAY DIFFRACTION4B40 - 289
5X-RAY DIFFRACTION5C8 - 160
6X-RAY DIFFRACTION6C161 - 289
7X-RAY DIFFRACTION7E5 - 137
8X-RAY DIFFRACTION8E138 - 288
9X-RAY DIFFRACTION9F6 - 41
10X-RAY DIFFRACTION10F42 - 289
11X-RAY DIFFRACTION11H5 - 133
12X-RAY DIFFRACTION12H134 - 289

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