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- PDB-5huo: Crystal Structure of NadC Deletion Mutant in C2221 Space Group -

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Basic information

Entry
Database: PDB / ID: 5huo
TitleCrystal Structure of NadC Deletion Mutant in C2221 Space Group
ComponentsNicotinate-nucleotide diphosphorylase (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
nicotinate-nucleotide diphosphorylase (carboxylating) / Quinolinate phosphoribosyltransferase [decarboxylating]
Similarity search - Component
Biological speciesStreptococcus pyogenes GA06023 (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.8 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 diphosphorylase (Carboxylating)
B: Nicotinate-nucleotide diphosphorylase (Carboxylating)
C: Nicotinate-nucleotide diphosphorylase (Carboxylating)
E: Nicotinate-nucleotide diphosphorylase (Carboxylating)
F: Nicotinate-nucleotide diphosphorylase (Carboxylating)
H: Nicotinate-nucleotide diphosphorylase (Carboxylating)
hetero molecules


Theoretical massNumber of molelcules
Total (without water)208,47726
Polymers206,5556
Non-polymers1,92120
Water2,684149
1
A: Nicotinate-nucleotide diphosphorylase (Carboxylating)
C: Nicotinate-nucleotide diphosphorylase (Carboxylating)
E: Nicotinate-nucleotide diphosphorylase (Carboxylating)
hetero molecules

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


Theoretical massNumber of molelcules
Total (without water)208,47726
Polymers206,5556
Non-polymers1,92120
Water1086
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation4_556x,-y,-z+11
Buried area29910 Å2
ΔGint-395 kcal/mol
Surface area58400 Å2
MethodPISA
2
B: Nicotinate-nucleotide diphosphorylase (Carboxylating)
F: Nicotinate-nucleotide diphosphorylase (Carboxylating)
H: Nicotinate-nucleotide diphosphorylase (Carboxylating)
hetero molecules

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


Theoretical massNumber of molelcules
Total (without water)208,47726
Polymers206,5556
Non-polymers1,92120
Water1086
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation3_655-x+1,y,-z+1/21
Buried area31130 Å2
ΔGint-403 kcal/mol
Surface area57680 Å2
MethodPISA
Unit cell
Length a, b, c (Å)106.203, 186.261, 221.906
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number20
Space group name H-MC2221
Components on special symmetry positions
IDModelComponents
11A-423-

HOH

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: _ / Beg auth comp-ID: THR / Beg label comp-ID: THR / Refine code: _

Dom-IDEns-IDEnd auth comp-IDEnd label comp-IDAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ASPASPAA6 - 28831 - 313
21ASPASPBB6 - 28831 - 313
12ASPASPAA6 - 28831 - 313
22ASPASPCC6 - 28831 - 313
13ASPASPAA6 - 28831 - 313
23ASPASPED6 - 28831 - 313
14LEULEUAA6 - 28731 - 312
24LEULEUFE6 - 28731 - 312
15ASPASPAA6 - 28831 - 313
25ASPASPHF6 - 28831 - 313
16ASPASPBB6 - 28831 - 313
26ASPASPCC6 - 28831 - 313
17ASPASPBB6 - 28831 - 313
27ASPASPED6 - 28831 - 313
18LEULEUBB6 - 28731 - 312
28LEULEUFE6 - 28731 - 312
19ASPASPBB6 - 28831 - 313
29ASPASPHF6 - 28831 - 313
110ASPASPCC6 - 28831 - 313
210ASPASPED6 - 28831 - 313
111ASPASPCC6 - 28831 - 313
211ASPASPFE6 - 28831 - 313
112ASPASPCC6 - 28831 - 313
212ASPASPHF6 - 28831 - 313
113ASPASPED6 - 28831 - 313
213ASPASPFE6 - 28831 - 313
114ASPASPED6 - 28831 - 313
214ASPASPHF6 - 28831 - 313
115ASPASPFE6 - 28831 - 313
215ASPASPHF6 - 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 diphosphorylase (Carboxylating)


Mass: 34425.902 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Streptococcus pyogenes GA06023 (bacteria)
Gene: nadC, HMPREF1231_0804 / Plasmid: pjExpress411 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3)
References: UniProt: A0A0H3BVM1, UniProt: U2UIT5*PLUS, nicotinate-nucleotide diphosphorylase (carboxylating)
#2: Chemical
ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 20 / Source method: obtained synthetically / Formula: SO4
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 149 / 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: 2.66 Å3/Da / Density % sol: 53.7 %
Crystal growTemperature: 293 K / Method: vapor diffusion / pH: 6.4
Details: 20% PEG 6000, 0.1 M BIS-TRIS, 0.48 M Ammonium sulfate, 1.25 mM Sodium Bromide

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-BM / Wavelength: 1 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Oct 17, 2014
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.8→40 Å / Num. obs: 54566 / % possible obs: 100 % / Observed criterion σ(I): -3 / Redundancy: 8.4 % / Rmerge(I) obs: 0.108 / Rsym value: 0.108 / Net I/σ(I): 14.6
Reflection shellResolution: 2.8→2.85 Å / Redundancy: 8.5 % / Rmerge(I) obs: 0.785 / Mean I/σ(I) obs: 2.3 / % possible all: 100

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Processing

Software
NameVersionClassification
REFMAC5.8.0135refinement
HKL-2000data reduction
HKL-2000data scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 5HUL
Resolution: 2.8→40 Å / Cor.coef. Fo:Fc: 0.925 / Cor.coef. Fo:Fc free: 0.892 / SU B: 33.335 / SU ML: 0.302 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.366 / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.25539 2766 5.1 %RANDOM
Rwork0.21599 ---
obs0.21799 51653 99.92 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.2 Å
Displacement parametersBiso mean: 124.789 Å2
Baniso -1Baniso -2Baniso -3
1-3.92 Å20 Å20 Å2
2---2.04 Å20 Å2
3----1.87 Å2
Refinement stepCycle: 1 / Resolution: 2.8→40 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms12807 0 100 149 13056
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0160.01913094
X-RAY DIFFRACTIONr_bond_other_d0.0090.0212388
X-RAY DIFFRACTIONr_angle_refined_deg1.5431.9617742
X-RAY DIFFRACTIONr_angle_other_deg2.206328329
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.73651689
X-RAY DIFFRACTIONr_dihedral_angle_2_deg30.06223.982570
X-RAY DIFFRACTIONr_dihedral_angle_3_deg15.971152153
X-RAY DIFFRACTIONr_dihedral_angle_4_deg22.2121583
X-RAY DIFFRACTIONr_chiral_restr0.0810.22111
X-RAY DIFFRACTIONr_gen_planes_refined0.0090.0214925
X-RAY DIFFRACTIONr_gen_planes_other0.0070.022994
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_it2.6352.3516771
X-RAY DIFFRACTIONr_mcbond_other2.6332.3516770
X-RAY DIFFRACTIONr_mcangle_it4.3963.518449
X-RAY DIFFRACTIONr_mcangle_other4.3963.518450
X-RAY DIFFRACTIONr_scbond_it2.7972.4146322
X-RAY DIFFRACTIONr_scbond_other2.6332.3246240
X-RAY DIFFRACTIONr_scangle_it
X-RAY DIFFRACTIONr_scangle_other4.3573.4579171
X-RAY DIFFRACTIONr_long_range_B_refined8.02617.98614308
X-RAY DIFFRACTIONr_long_range_B_other8.01717.96914300
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)
11A334760.1
12B334760.1
21A286580.14
22C286580.14
31A319140.11
32E319140.11
41A336800.08
42F336800.08
51A324760.1
52H324760.1
61B289440.13
62C289440.13
71B325460.1
72E325460.1
81B334480.09
82F334480.09
91B327460.09
92H327460.09
101C273660.15
102E273660.15
111C284060.14
112F284060.14
121C288660.14
122H288660.14
131E322620.1
132F322620.1
141E312040.11
142H312040.11
151F324500.1
152H324500.1
LS refinement shellResolution: 2.8→2.872 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.324 222 -
Rwork0.317 3736 -
obs--100 %
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)
10.8432-0.1738-0.01193.64592.18121.75390.121-0.0627-0.1989-0.3993-0.24820.0407-0.2872-0.68870.12720.54020.172-0.0240.6966-0.0140.353635.073212.587104.3724
20.28050.34390.17590.6836-0.07680.5097-0.0610.0278-0.12430.00110.1039-0.0527-0.1233-0.1128-0.04290.5630.01930.00510.51570.03520.42447.08483.098198.0728
30.960.58210.39381.8183-0.04720.58860.3133-0.177-0.42230.3827-0.2447-0.63690.0487-0.0007-0.06860.5231-0.0446-0.12660.40210.10460.522352.7039-20.3562104.3663
41.56023.14860.74668.19730.43171.00910.1003-0.25470.0740.3698-0.4064-0.0889-0.0531-0.17710.3060.48140.05250.00920.5057-0.00110.630742.45641.450762.1519
50.21170.1806-0.51880.8940.90483.77490.0096-0.0627-0.11830.1192-0.3843-0.0449-0.0141-0.48380.37470.36960.10040.01670.5838-0.1290.515228.565943.44665.8864
60.06030.07490.2510.89970.29971.2095-0.0083-0.10520.023-0.19530.01030.1378-0.0804-0.2256-0.00190.4040.0368-0.05350.53-0.05940.561826.147530.311742.1388
71.15780.9435-0.42571.05290.21381.46340.0488-0.08550.4395-0.0564-0.23640.4133-0.0045-0.26710.18770.3682-0.0239-0.00650.5519-0.10590.673713.1199-8.944592.3171
813.9971-14.29960.899320.59521.29540.8794-0.3072-0.0375-0.2517-0.01450.4360.2989-0.08120.1856-0.12880.5041-0.0966-0.110.6537-0.21930.6554-8.2925-14.8552120.3125
90.6810.4701-0.70340.77240.19862.3123-0.2407-0.12390.1703-0.065-0.54480.42210.5359-0.62170.78560.2397-0.27790.13651.049-0.59720.7501-8.5188-14.6097105.2117
107.0214-2.1174-2.25781.5751.54851.5840.55760.24210.64130.433-0.64070.00820.4072-0.53090.08310.4431-0.32220.24670.7021-0.18410.490911.7321-10.5736118.1808
110.41960.43670.37630.6520.47170.37060.3488-0.20210.12530.2664-0.4290.03340.2626-0.26210.08020.6191-0.32510.11890.6706-0.07770.382317.1304-24.6171108.9221
121.59351.1415-1.18651.0511-0.30982.3840.08340.20180.06890.16310.034-0.05510.433-0.2817-0.11740.5768-0.0847-0.04730.46350.01780.505828.0327-30.114591.1135
131.3255-0.8341.22011.215-0.19551.663-0.07850.00970.19-0.0698-0.03530.0005-0.21190.05160.11380.4879-0.02120.01490.53240.03690.474765.471618.248348.6569
140.7304-0.19620.00690.2713-0.00260.02330.0285-0.00360.0836-0.04980.01840.07090.0950.0152-0.04690.5601-0.0313-0.00690.51050.03070.431845.56925.672345.5479
153.3770.2739-0.21750.639-0.0240.51460.0142-0.2317-0.1134-0.01510.00950.12920.25530.0098-0.02370.5411-0.0369-0.02970.43490.05720.392443.1252-2.979345.5229
160.38020.40590.24592.35461.23460.67510.04350.0220.0457-0.11990.1636-0.2870.00510.0298-0.2070.48030.0330.03020.5077-0.02560.546447.134828.866941.2481
170.0874-0.22770.05741.80370.83240.84760.0452-0.02770.0328-0.1174-0.1001-0.0868-0.0216-0.11220.05490.51690.0650.02730.426-0.02510.575843.748242.836335.6445
180.8208-0.42520.28570.80220.46630.805-0.027-0.33610.067-0.056-0.1539-0.0151-0.129-0.41570.1810.4850.052-0.02080.4803-0.02350.459138.952462.104751.4997
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A6 - 39
2X-RAY DIFFRACTION2A40 - 168
3X-RAY DIFFRACTION3A169 - 288
4X-RAY DIFFRACTION4B6 - 36
5X-RAY DIFFRACTION5B37 - 84
6X-RAY DIFFRACTION6B85 - 288
7X-RAY DIFFRACTION7C6 - 169
8X-RAY DIFFRACTION8C170 - 177
9X-RAY DIFFRACTION9C181 - 288
10X-RAY DIFFRACTION10E6 - 39
11X-RAY DIFFRACTION11E40 - 182
12X-RAY DIFFRACTION12E183 - 288
13X-RAY DIFFRACTION13F5 - 39
14X-RAY DIFFRACTION14F40 - 232
15X-RAY DIFFRACTION15F233 - 288
16X-RAY DIFFRACTION16H6 - 36
17X-RAY DIFFRACTION17H37 - 168
18X-RAY DIFFRACTION18H169 - 288

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