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- PDB-6gg9: Crystal structures of a short blue light photoreceptor protein Pp... -

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Basic information

Entry
Database: PDB / ID: 6gg9
TitleCrystal structures of a short blue light photoreceptor protein PpSB1-LOV mutant (dark state) - R61H/R66I
ComponentsSensory box protein
KeywordsSIGNALING PROTEIN / blue light photoreceptor / Sensory box protein
Function / homology
Function and homology information


PAS domain / PAS-associated, C-terminal / PAC domain profile. / PAC motif / Motif C-terminal to PAS motifs (likely to contribute to PAS structural domain) / PAS domain / Beta-Lactamase / PAS domain / PAS repeat profile. / PAS domain ...PAS domain / PAS-associated, C-terminal / PAC domain profile. / PAC motif / Motif C-terminal to PAS motifs (likely to contribute to PAS structural domain) / PAS domain / Beta-Lactamase / PAS domain / PAS repeat profile. / PAS domain / PAS domain superfamily / 2-Layer Sandwich / Alpha Beta
Similarity search - Domain/homology
FLAVIN MONONUCLEOTIDE / Sensory box protein
Similarity search - Component
Biological speciesPseudomonas putida (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.04 Å
AuthorsRoellen, K. / Granzin, J. / Batra-Safferling, R.
CitationJournal: PLoS One / Year: 2018
Title: Small-angle X-ray scattering study of the kinetics of light-dark transition in a LOV protein.
Authors: Katrin Röllen / Joachim Granzin / Renu Batra-Safferling / Andreas Maximilian Stadler /
Abstract: Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue- ...Light, oxygen, voltage (LOV) photoreceptors consist of conserved photo-responsive domains in bacteria, archaea, plants and fungi, and detect blue-light via a flavin cofactor. We investigated the blue-light induced conformational transition of the dimeric photoreceptor PpSB1-LOV-R66I from Pseudomonas putida in solution by using small-angle X-ray scattering (SAXS). SAXS experiments of the fully populated light- and dark-states under steady-state conditions revealed significant structural differences between the two states that are in agreement with the known structures determined by crystallography. We followed the transition from the light- to the dark-state by using SAXS measurements in real-time. A two-state model based on the light- and dark-state conformations could describe the measured time-course SAXS data with a relaxation time τREC of ~ 34 to 35 min being larger than the recovery time found with UV/vis spectroscopy. Unlike the flavin chromophore-based UV/vis method that is sensitive to the local chromophore environment in flavoproteins, SAXS-based assay depends on protein conformational changes and provides with an alternative to measure the recovery kinetics.
History
DepositionMay 3, 2018Deposition site: PDBE / Processing site: PDBE
Revision 1.0Jul 25, 2018Provider: repository / Type: Initial release
Revision 1.1Jan 17, 2024Group: 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
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Sensory box protein
B: Sensory box protein
C: Sensory box protein
D: Sensory box protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)76,0448
Polymers74,2194
Non-polymers1,8254
Water1,982110
1
A: Sensory box protein
C: Sensory box protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)38,0224
Polymers37,1092
Non-polymers9132
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4850 Å2
ΔGint-38 kcal/mol
Surface area13500 Å2
MethodPISA
2
B: Sensory box protein
D: Sensory box protein
hetero molecules


Theoretical massNumber of molelcules
Total (without water)38,0224
Polymers37,1092
Non-polymers9132
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4910 Å2
ΔGint-39 kcal/mol
Surface area13660 Å2
MethodPISA
Unit cell
Length a, b, c (Å)102.700, 76.506, 83.279
Angle α, β, γ (deg.)90.00, 106.23, 90.00
Int Tables number5
Space group name H-MC121

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Components

#1: Protein
Sensory box protein


Mass: 18554.746 Da / Num. of mol.: 4 / Mutation: R61H, R66I
Source method: isolated from a genetically manipulated source
Details: expression tag: MGSSHHHHHHSSGLVPRGSH / Source: (gene. exp.) Pseudomonas putida (bacteria) / Gene: PP_4629
Production host: Escherichia coli 'BL21-Gold(DE3)pLysS AG' (bacteria)
References: UniProt: Q88E39
#2: Chemical
ChemComp-FMN / FLAVIN MONONUCLEOTIDE / RIBOFLAVIN MONOPHOSPHATE


Mass: 456.344 Da / Num. of mol.: 4 / Source method: obtained synthetically / Formula: C17H21N4O9P
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 110 / 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: 2.12 Å3/Da / Density % sol: 42.12 %
Crystal growTemperature: 292.15 K / Method: vapor diffusion, sitting drop / pH: 6.6 / Details: 0.2 MgCl2, 0.1M MES, 15% (w/v) PEG4000

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: ESRF / Beamline: ID29 / Wavelength: 0.9772 Å
DetectorType: DECTRIS PILATUS 6M-F / Detector: PIXEL / Date: Jun 20, 2014
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9772 Å / Relative weight: 1
ReflectionResolution: 2.04→48.45 Å / Num. obs: 39334 / % possible obs: 99.5 % / Redundancy: 4.4 % / Biso Wilson estimate: 37.5 Å2 / Rmerge(I) obs: 0.069 / Rrim(I) all: 0.078 / Net I/σ(I): 11.2
Reflection shellResolution: 2.04→2.1 Å / Redundancy: 4.4 % / Mean I/σ(I) obs: 1.7 / Num. unique obs: 3076 / % possible all: 99.7

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Processing

Software
NameVersionClassification
PHENIX(1.10.1-2155_1692: ???)refinement
XDSdata reduction
Aimlessdata scaling
MOLREPphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 5J3W
Resolution: 2.04→48.45 Å / SU ML: 0.29 / Cross valid method: FREE R-VALUE / σ(F): 1.37 / Phase error: 28.27
RfactorNum. reflection% reflection
Rfree0.2367 1955 4.98 %
Rwork0.1947 --
obs0.1968 39265 99.27 %
Solvent computationShrinkage radii: 1 Å / VDW probe radii: 1.2 Å
Refinement stepCycle: LAST / Resolution: 2.04→48.45 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4165 0 124 110 4399
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0084373
X-RAY DIFFRACTIONf_angle_d0.9285941
X-RAY DIFFRACTIONf_dihedral_angle_d14.7132596
X-RAY DIFFRACTIONf_chiral_restr0.055652
X-RAY DIFFRACTIONf_plane_restr0.006770
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.04-2.0910.35811560.30362648X-RAY DIFFRACTION99
2.091-2.14760.35881470.29622629X-RAY DIFFRACTION99
2.1476-2.21080.30411260.2822661X-RAY DIFFRACTION99
2.2108-2.28210.31071310.26122624X-RAY DIFFRACTION98
2.2821-2.36370.30811480.23362665X-RAY DIFFRACTION100
2.3637-2.45830.28471280.23942675X-RAY DIFFRACTION99
2.4583-2.57020.25961280.22532673X-RAY DIFFRACTION99
2.5702-2.70570.27511240.21992631X-RAY DIFFRACTION99
2.7057-2.87520.26961490.23082645X-RAY DIFFRACTION99
2.8752-3.09710.2741480.23022677X-RAY DIFFRACTION100
3.0971-3.40870.25841490.19982675X-RAY DIFFRACTION100
3.4087-3.90180.19721440.17192662X-RAY DIFFRACTION99
3.9018-4.91510.18521380.14312712X-RAY DIFFRACTION100
4.9151-48.46360.19481390.16352733X-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)
10.4892-0.1412-0.46151.99580.99790.7204-0.35950.23190.2283-0.23140.56880.7074-0.9802-0.89650.00010.46860.0602-0.02760.55270.07420.5076-9.38951.044320.5138
21.3144-0.79760.75680.96640.25451.30140.3732-0.2080.31790.2296-0.4029-0.1389-0.38960.59650.00160.393-0.0093-0.02030.38750.06370.36698.5616-2.720123.2945
30.02990.07340.08590.5211-0.04450.53640.2327-0.5280.51950.29910.0012-0.6386-0.06361.28350.00080.46780.0263-0.06130.56950.0340.469414.71650.652822.6957
42.00770.1426-0.6250.2514-0.20430.29450.0478-1.61710.28310.8868-0.5160.2702-0.9956-0.0707-0.01240.4611-0.02040.02490.3944-0.04710.37375.5046-3.699833.923
50.86030.053-0.02150.7684-0.24781.46520.2696-0.83160.20240.2485-0.5033-0.1433-1.09991.0038-0.01360.6224-0.1051-0.03610.5803-0.0670.499414.23850.215630.895
60.00560.01320.00290.12420.0612-0.00070.0189-0.0245-0.5176-0.31550.0172-0.8962-1.0760.714-0.00230.91230.0681-0.15541.3573-0.00571.039823.33-13.312431.5045
70.2710.74110.26222.04870.64930.5239-0.63811.733-0.7765-1.09991.0084-1.34840.82691.21290.0270.4964-0.00190.12561.1056-0.20430.883119.4298-13.658119.0786
84.47320.2591-0.5543.2702-0.30571.36340.0167-0.0468-0.64090.0476-0.0091-0.280.31060.6229-0.00040.43120.0575-0.02940.5039-0.00130.412512.1216-11.649526.0168
91.77590.14940.44030.77740.67040.6927-0.15130.59450.11280.30610.50430.103-0.3762-0.22480.0090.3384-0.0116-0.03690.4290.12570.28328.299-3.007817.6651
100.39960.0247-0.7114-0.0033-0.08231.2607-0.0216-1.312-1.69061.54880.6752-1.29530.93060.68970.47420.94320.0702-0.17480.5580.39850.99826.9201-27.401338.3391
111.20371.55671.05922.00421.36360.9353-0.0491.0836-0.03580.56290.6146-2.0501-0.72041.96110.11730.49140.08260.0010.6936-0.00040.504817.1207-7.853124.4088
121.0273-0.78260.32880.5839-0.4110.68920.25751.03380.31430.1868-0.08840.6373-0.007-1.18620.00010.41850.0297-0.00370.6144-0.04720.492615.6201-29.98470.1889
130.97770.12770.8153.11381.07421.44620.43530.1110.948-0.3840.1513-0.2509-0.63280.3299-0.00020.43740.01510.01370.4594-0.07290.476531.313-22.98712.5827
140.45970.0075-0.46081.2380.49190.7440.6844-0.20560.976-0.83720.2833-0.6039-1.05740.56830.01520.75970.03780.15620.6137-0.09260.89735.1433-16.84891.9143
150.1294-0.3709-0.42362.40490.51081.36330.65420.08191.0752-0.846-1.0117-0.8748-0.68450.7388-0.00090.560.0845-0.00791.163-0.16010.875945.6582-27.36899.9729
164.931-0.804-1.9711.8861.13074.6698-0.1547-0.69780.3370.14570.3132-0.47520.13040.62690.00370.42220.1424-0.0240.5904-0.10210.419335.1604-27.88587.0836
170.41210.23450.13821.0634-0.38670.46450.405-0.66710.10940.1887-0.0327-0.4736-0.1981.2099-0.03070.94650.255-0.24651.0486-0.17240.6933.7602-21.06832.6902
180.0938-0.0050.030.09860.09060.09161.0852-1.16360.9164-0.44330.4482-2.5148-0.89421.1735-0.0020.6719-0.00780.22720.8903-0.33611.001140.6306-25.01245.8478
192.1893-0.17851.06681.8833-0.06183.1209-0.22360.53730.1749-0.23060.07960.10480.033-0.1071-0.00040.36520.0263-0.01940.43230.06650.3848-5.018-3.610619.5445
201.17280.85610.70441.0237-0.12131.3404-0.61540.6041-0.72570.13480.42010.7330.1354-0.5381-0.0010.44570.00320.03640.60770.00660.5125-13.9391-11.404520.261
210.1146-0.1357-0.04641.03880.51360.2345-0.32860.6988-0.2621-0.30910.50241.3377-0.2203-0.4931-0.0010.691-0.107-0.02561.0165-0.00510.7667-18.2108-14.511218.4213
220.02710.04240.05660.03280.07330.105-0.18710.31250.1032-0.4604-0.28060.7263-0.1105-2.2025-0.00351.1341-0.34650.05911.2037-0.04060.9114-22.5364-22.247731.9429
234.34631.6776-1.07352.5316-1.55536.7032-0.0695-0.06510.15430.6253-0.00080.4467-0.0654-0.50820.01080.4078-0.00110.06650.4125-0.05340.5231-14.5267-10.540530.7254
240.9237-0.2031-0.50440.1508-0.0410.8397-0.2328-1.7232-0.91780.33090.2448-0.10280.278-0.47540.04460.97130.14710.09720.76760.14531.1392-3.2445-29.335639.3276
251.2006-0.66213.15143.2224-0.49768.78540.37290.1062-0.51970.0393-0.33690.85361.065-1.05420.50290.44760.0405-0.00220.0964-0.07430.389-19.3024-12.219728.8206
260.6654-0.28370.16450.9850.74460.7605-0.23330.7346-0.0854-0.3581-0.1652-0.20770.09260.1643-0.00050.43250.03730.07370.5917-0.09720.537324.2987-33.4517-1.0039
271.19120.3603-0.25833.5153-0.99712.5399-0.1845-0.3982-0.6493-0.02670.04030.01430.3263-0.28990.00260.45190.07140.03970.4574-0.0180.436712.4069-36.470211.3888
280.51860.12010.2140.6375-0.40220.38510.0153-0.454-1.0794-0.0084-0.17050.27930.7571-0.4458-0.0020.60910.03010.07630.52420.00140.7199.4214-41.919315.6494
292.22341.3527-0.38262.65191.13411.0270.223-0.73560.43410.76080.03491.1904-0.2934-0.22750.0010.5490.09670.1470.6414-0.06570.76896.3583-27.509821.4697
302.48420.2239-0.60131.79351.29431.4315-0.2749-0.4426-0.21450.42550.14840.0417-0.1661-0.2903-0.00020.33240.0856-0.00980.44570.03220.452813.9056-32.410717.8032
310.79250.0447-0.08040.7281-0.16480.32940.18890.5094-0.05610.03260.16460.10840.10230.24970.00030.37510.0288-0.03790.4584-0.03090.50249.7879-30.24386.1709
320.67640.1137-0.63430.85310.52520.9798-0.6357-1.480.6802-0.48240.5769-0.9373-1.8044-0.0790.05451.06270.011-0.23991.01110.04580.48424.3135-22.368834.9442
330.4481-0.05770.07490.0878-0.07970.0622-0.1064-0.8805-0.50740.4538-0.27081.29110.9591-0.69760.00580.45430.01120.05320.5279-0.02610.46046.2295-32.236917.5746
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid -1 through 13 )
2X-RAY DIFFRACTION2chain 'A' and (resid 14 through 24 )
3X-RAY DIFFRACTION3chain 'A' and (resid 25 through 33 )
4X-RAY DIFFRACTION4chain 'A' and (resid 34 through 41 )
5X-RAY DIFFRACTION5chain 'A' and (resid 42 through 57 )
6X-RAY DIFFRACTION6chain 'A' and (resid 58 through 63 )
7X-RAY DIFFRACTION7chain 'A' and (resid 64 through 74 )
8X-RAY DIFFRACTION8chain 'A' and (resid 75 through 104 )
9X-RAY DIFFRACTION9chain 'A' and (resid 105 through 119 )
10X-RAY DIFFRACTION10chain 'A' and (resid 120 through 132 )
11X-RAY DIFFRACTION11chain 'A' and (resid 500 through 500 )
12X-RAY DIFFRACTION12chain 'B' and (resid 1 through 12 )
13X-RAY DIFFRACTION13chain 'B' and (resid 13 through 41 )
14X-RAY DIFFRACTION14chain 'B' and (resid 42 through 57 )
15X-RAY DIFFRACTION15chain 'B' and (resid 58 through 74 )
16X-RAY DIFFRACTION16chain 'B' and (resid 75 through 119 )
17X-RAY DIFFRACTION17chain 'B' and (resid 120 through 135 )
18X-RAY DIFFRACTION18chain 'B' and (resid 500 through 500 )
19X-RAY DIFFRACTION19chain 'C' and (resid 1 through 24 )
20X-RAY DIFFRACTION20chain 'C' and (resid 25 through 41 )
21X-RAY DIFFRACTION21chain 'C' and (resid 42 through 57 )
22X-RAY DIFFRACTION22chain 'C' and (resid 58 through 63 )
23X-RAY DIFFRACTION23chain 'C' and (resid 64 through 119 )
24X-RAY DIFFRACTION24chain 'C' and (resid 120 through 134 )
25X-RAY DIFFRACTION25chain 'C' and (resid 500 through 500 )
26X-RAY DIFFRACTION26chain 'D' and (resid 1 through 13 )
27X-RAY DIFFRACTION27chain 'D' and (resid 14 through 41 )
28X-RAY DIFFRACTION28chain 'D' and (resid 42 through 57 )
29X-RAY DIFFRACTION29chain 'D' and (resid 58 through 86 )
30X-RAY DIFFRACTION30chain 'D' and (resid 87 through 104 )
31X-RAY DIFFRACTION31chain 'D' and (resid 105 through 119 )
32X-RAY DIFFRACTION32chain 'D' and (resid 120 through 135 )
33X-RAY DIFFRACTION33chain 'D' and (resid 500 through 500 )

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