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- PDB-1r3n: Crystal structure of beta-alanine synthase from Saccharomyces kluyveri -

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Basic information

Entry
Database: PDB / ID: 1r3n
TitleCrystal structure of beta-alanine synthase from Saccharomyces kluyveri
Componentsbeta-alanine synthase
KeywordsHYDROLASE / alpha and beta protein / di-zinc center
Function / homology
Function and homology information


beta-ureidopropionase / beta-ureidopropionase activity / hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in linear amidines / metal ion binding
Similarity search - Function
Amidase, carbamoylase-type / Alpha-Beta Plaits - #360 / Bacterial exopeptidase dimerisation domain / Peptidase M20, dimerisation domain / Peptidase dimerisation domain / Peptidase M20 / Peptidase family M20/M25/M40 / Zn peptidases / Aminopeptidase / Alpha-Beta Plaits ...Amidase, carbamoylase-type / Alpha-Beta Plaits - #360 / Bacterial exopeptidase dimerisation domain / Peptidase M20, dimerisation domain / Peptidase dimerisation domain / Peptidase M20 / Peptidase family M20/M25/M40 / Zn peptidases / Aminopeptidase / Alpha-Beta Plaits / 2-Layer Sandwich / 3-Layer(aba) Sandwich / Alpha Beta
Similarity search - Domain/homology
BETA-AMINO ISOBUTYRATE / Beta-alanine synthase
Similarity search - Component
Biological speciesLachancea kluyveri (fungus)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.7 Å
AuthorsLundgren, S. / Gojkovic, Z. / Piskur, J. / Dobritzsch, D.
Citation
Journal: J.Biol.Chem. / Year: 2003
Title: Yeast beta-Alanine Synthase Shares a Structural Scaffold and Origin with Dizinc-dependent Exopeptidases
Authors: Lundgren, S. / Gojkovic, Z. / Piskur, J. / Dobritzsch, D.
#1: Journal: Acta Crystallogr.,Sect.D / Year: 2003
Title: Crystallization and preliminary X-ray analysis of beta-alanine synthase from the yeast Saccharomyces kluyveri
Authors: Dobritzsch, D. / Gojkovic, Z. / Andersen, B. / Piskur, J.
History
DepositionOct 2, 2003Deposition site: RCSB / Processing site: RCSB
Revision 1.0Nov 11, 2003Provider: repository / Type: Initial release
Revision 1.1Apr 29, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3Feb 14, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_struct_conn_angle / struct_conn / struct_ncs_dom_lim / struct_ref_seq_dif / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_struct_conn_angle.ptnr1_auth_comp_id / _pdbx_struct_conn_angle.ptnr1_auth_seq_id / _pdbx_struct_conn_angle.ptnr1_label_asym_id / _pdbx_struct_conn_angle.ptnr1_label_atom_id / _pdbx_struct_conn_angle.ptnr1_label_comp_id / _pdbx_struct_conn_angle.ptnr1_label_seq_id / _pdbx_struct_conn_angle.ptnr3_auth_comp_id / _pdbx_struct_conn_angle.ptnr3_auth_seq_id / _pdbx_struct_conn_angle.ptnr3_label_asym_id / _pdbx_struct_conn_angle.ptnr3_label_atom_id / _pdbx_struct_conn_angle.ptnr3_label_comp_id / _pdbx_struct_conn_angle.ptnr3_label_seq_id / _pdbx_struct_conn_angle.value / _struct_conn.pdbx_dist_value / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_label_seq_id / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_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_seq_id / _struct_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id
Revision 1.4Apr 3, 2024Group: Refinement description / Category: pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: beta-alanine synthase
B: beta-alanine synthase
C: beta-alanine synthase
D: beta-alanine synthase
E: beta-alanine synthase
F: beta-alanine synthase
G: beta-alanine synthase
H: beta-alanine synthase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)407,69232
Polymers405,8298
Non-polymers1,86324
Water9,170509
1
A: beta-alanine synthase
B: beta-alanine synthase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)101,9238
Polymers101,4572
Non-polymers4666
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5740 Å2
ΔGint-179 kcal/mol
Surface area33160 Å2
MethodPISA
2
C: beta-alanine synthase
D: beta-alanine synthase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)101,9238
Polymers101,4572
Non-polymers4666
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5810 Å2
ΔGint-184 kcal/mol
Surface area33370 Å2
MethodPISA
3
E: beta-alanine synthase
F: beta-alanine synthase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)101,9238
Polymers101,4572
Non-polymers4666
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5820 Å2
ΔGint-184 kcal/mol
Surface area33620 Å2
MethodPISA
4
G: beta-alanine synthase
H: beta-alanine synthase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)101,9238
Polymers101,4572
Non-polymers4666
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5740 Å2
ΔGint-181 kcal/mol
Surface area33420 Å2
MethodPISA
Unit cell
Length a, b, c (Å)117.230, 77.120, 225.520
Angle α, β, γ (deg.)90.00, 95.05, 90.00
Int Tables number4
Space group name H-MP1211
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B
31C
41D
51E
61F
71G
81H
12A
22B
32C
42D
52F
62H
72E
82G
92A
102B
112C
122D
132F
142H
152E
162G
172A
182B
192C
202D
212F
222H
232E
242G
252A
262B
272C
282D
292F
302H
312E
322G
332A
342B
352C
362D
372F
382H
392E
402G

NCS domain segments:
Dom-IDComponent-IDEns-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
111ALAALAPROPRO2AA248 - 363247 - 362
211ALAALAPROPRO2BB248 - 363247 - 362
311ALAALAPROPRO2CC248 - 363247 - 362
411ALAALAPROPRO2DD248 - 363247 - 362
511ALAALAPROPRO2EE248 - 363247 - 362
611ALAALAPROPRO2FF248 - 363247 - 362
711ALAALAPROPRO2GG248 - 363247 - 362
811ALAALAPROPRO2HH248 - 363247 - 362
112ALAALALEULEU3AA31 - 18630 - 185
212ALAALALEULEU3BB31 - 18630 - 185
312ALAALALEULEU3CC31 - 18630 - 185
412ALAALALEULEU3DD31 - 18630 - 185
512ALAALALEULEU3FF31 - 18630 - 185
612ALAALALEULEU3HH31 - 18630 - 185
712ALAALALEULEU3EE31 - 18630 - 185
812ALAALALEULEU3GG31 - 18630 - 185
922VALVALVALVAL3AA197 - 246196 - 245
1022VALVALVALVAL3BB197 - 246196 - 245
1122VALVALVALVAL3CC197 - 246196 - 245
1222VALVALVALVAL3DD197 - 246196 - 245
1322VALVALVALVAL3FF197 - 246196 - 245
1422VALVALVALVAL3HH197 - 246196 - 245
1522VALVALVALVAL3EE197 - 246196 - 245
1622VALVALVALVAL3GG197 - 246196 - 245
1732VALVALTYRTYR3AA365 - 423364 - 422
1832VALVALTYRTYR3BB365 - 423364 - 422
1932VALVALTYRTYR3CC365 - 423364 - 422
2032VALVALTYRTYR3DD365 - 423364 - 422
2132VALVALTYRTYR3FF365 - 423364 - 422
2232VALVALTYRTYR3HH365 - 423364 - 422
2332VALVALTYRTYR3EE365 - 423364 - 422
2432VALVALTYRTYR3GG365 - 423364 - 422
2542GLUGLUILEILE3AA431 - 452430 - 451
2642GLUGLUILEILE3BB431 - 452430 - 451
2742GLUGLUILEILE3CC431 - 452430 - 451
2842GLUGLUILEILE3DD431 - 452430 - 451
2942GLUGLUILEILE3FF431 - 452430 - 451
3042GLUGLUILEILE3HH431 - 452430 - 451
3142GLUGLUILEILE3EE431 - 452430 - 451
3242GLUGLUILEILE3GG431 - 452430 - 451
3352ZNZNZNZN1AI - J500 - 501
3452ZNZNZNZN1BL - M500 - 501
3552ZNZNZNZN1CO - P500 - 501
3652ZNZNZNZN1DR - S500 - 501
3752ZNZNZNZN1FY - Z500 - 501
3852ZNZNZNZN1HEA - FA500 - 501
3952ZNZNZNZN1EU - V500 - 501
4052ZNZNZNZN1GAA - BA500 - 501

NCS ensembles :
ID
1
2
DetailsThe biological assembly is a dimer. The asymmetric unit contains four such dimers.

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Components

#1: Protein
beta-alanine synthase


Mass: 50728.598 Da / Num. of mol.: 8
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Lachancea kluyveri (fungus) / Gene: PYD3 / Plasmid: P491 / Production host: Escherichia coli (E. coli) / Strain (production host): BL21-CodonPlus / References: UniProt: Q96W94, beta-ureidopropionase
#2: Chemical
ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 16 / Source method: obtained synthetically / Formula: Zn
#3: Chemical
ChemComp-BIB / BETA-AMINO ISOBUTYRATE


Mass: 102.112 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: C4H8NO2
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 509 / 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: 2.5 Å3/Da / Density % sol: 50.82 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 6
Details: trisodium citrate, dioxane, pH 6.0, VAPOR DIFFUSION, HANGING DROP, temperature 293K
Crystal grow
*PLUS
Temperature: 20 ℃ / pH: 7.5 / Method: vapor diffusion, hanging drop
Components of the solutions
*PLUS
IDConc.Common nameCrystal-IDSol-IDDetailsChemical formula
14.0-4.5 mg/mlprotein1drop
250 mMTris1droppH7.5
31 mMdithiothreitol1drop
4100 mM1dropNaCl
50.88 Mtrisodium citrate1reservoir
60.1 Msodium citrate1reservoirpH6.0
75 %(v/v)dioxane1reservoir

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID14-1 / Wavelength: 0.933 Å
DetectorType: ADSC QUANTUM 4 / Detector: CCD / Date: Dec 10, 2001 / Details: diamond monochromator and bent multilayer
RadiationMonochromator: diamond / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.933 Å / Relative weight: 1
ReflectionResolution: 2.7→25 Å / Num. obs: 106190 / % possible obs: 95.9 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 3.5 % / Biso Wilson estimate: 54.6 Å2 / Rmerge(I) obs: 0.083 / Net I/σ(I): 14.8
Reflection shellResolution: 2.7→2.85 Å / Redundancy: 2.6 % / Rmerge(I) obs: 0.261 / Mean I/σ(I) obs: 3.7 / Num. unique all: 13170 / % possible all: 82.2
Reflection shell
*PLUS
% possible obs: 82.2 %

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Processing

Software
NameVersionClassification
REFMAC5.1.24refinement
MOSFLMdata reduction
CCP4(SCALA)data scaling
EPMRphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: model of seleno-methionine substituted beta-alanine synthase from the same source

Resolution: 2.7→25 Å / Cor.coef. Fo:Fc: 0.944 / Cor.coef. Fo:Fc free: 0.904 / SU B: 15.258 / SU ML: 0.301 / Isotropic thermal model: Isotropic / Cross valid method: THROUGHOUT / σ(F): 0 / σ(I): 0 / ESU R Free: 0.387 / Stereochemistry target values: MAXIMUM LIKELIHOOD
RfactorNum. reflection% reflectionSelection details
Rfree0.26577 5296 5 %RANDOM
Rwork0.20837 ---
obs0.21118 100879 95.84 %-
all-100879 --
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 16.339 Å2
Baniso -1Baniso -2Baniso -3
1-1.21 Å20 Å22.2 Å2
2---2.46 Å20 Å2
3---1.63 Å2
Refinement stepCycle: LAST / Resolution: 2.7→25 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms26906 0 72 519 27497
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0090.02127622
X-RAY DIFFRACTIONr_angle_refined_deg1.1891.93837391
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.30153477
X-RAY DIFFRACTIONr_chiral_restr0.0890.24048
X-RAY DIFFRACTIONr_gen_planes_refined0.0040.0221260
X-RAY DIFFRACTIONr_nbd_refined0.2050.212807
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.1640.2
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2220.2143
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.2180.220
X-RAY DIFFRACTIONr_mcbond_it0.1851.517236
X-RAY DIFFRACTIONr_mcangle_it0.447227646
X-RAY DIFFRACTIONr_scbond_it0.97310386
X-RAY DIFFRACTIONr_scangle_it1.6214.59745
Refine LS restraints NCS

Refine-ID: X-RAY DIFFRACTION

Ens-IDDom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
11A464tight positional0.040.05
12B464tight positional0.040.05
13C464tight positional0.040.05
14D464tight positional0.040.05
15E464tight positional0.030.05
16F464tight positional0.040.05
17G464tight positional0.030.05
18H464tight positional0.030.05
21A1150tight positional0.070.05
22B1150tight positional0.030.05
23C1150tight positional0.040.05
24D1150tight positional0.030.05
25F1150tight positional0.040.05
26H1150tight positional0.030.05
27E1150tight positional0.030.05
28G1150tight positional0.030.05
11A427medium positional0.470.5
12B427medium positional0.490.5
13C427medium positional0.470.5
14D427medium positional0.460.5
15E427medium positional0.470.5
16F427medium positional0.460.5
17G427medium positional0.480.5
18H427medium positional0.50.5
21A1081loose positional0.585
22B1081loose positional0.715
23C1081loose positional0.565
24D1081loose positional0.75
25F1081loose positional0.595
26H1081loose positional0.645
27E1081loose positional1.075
28G1081loose positional0.85
11A464tight thermal0.080.5
12B464tight thermal0.060.5
13C464tight thermal0.060.5
14D464tight thermal0.050.5
15E464tight thermal0.060.5
16F464tight thermal0.050.5
17G464tight thermal0.050.5
18H464tight thermal0.050.5
21A1150tight thermal0.070.5
22B1150tight thermal0.050.5
23C1150tight thermal0.060.5
24D1150tight thermal0.050.5
25F1150tight thermal0.070.5
26H1150tight thermal0.060.5
27E1150tight thermal0.060.5
28G1150tight thermal0.060.5
11A427medium thermal0.662
12B427medium thermal0.452
13C427medium thermal0.52
14D427medium thermal0.452
15E427medium thermal0.442
16F427medium thermal0.472
17G427medium thermal0.482
18H427medium thermal0.462
21A1081loose thermal1.5110
22B1081loose thermal1.110
23C1081loose thermal1.5410
24D1081loose thermal1.0810
25F1081loose thermal1.3610
26H1081loose thermal1.1510
27E1081loose thermal1.4510
28G1081loose thermal1.5310
LS refinement shellResolution: 2.7→2.77 Å / Total num. of bins used: 20
RfactorNum. reflection
Rfree0.355 318
Rwork0.277 6251
obs-6632
Refinement
*PLUS
Highest resolution: 2.7 Å / Lowest resolution: 30 Å / Rfactor Rfree: 0.266 / Rfactor Rwork: 0.208
Solvent computation
*PLUS
Displacement parameters
*PLUS
Refine LS restraints
*PLUS
Refine-IDTypeDev ideal
X-RAY DIFFRACTIONr_bond_d0.009
X-RAY DIFFRACTIONr_angle_d
X-RAY DIFFRACTIONr_angle_deg1.2

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