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- PDB-9j24: Structural basis of the bifunctionality of M. salinexigens ZYF650... -

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Basic information

Entry
Database: PDB / ID: 9j24
TitleStructural basis of the bifunctionality of M. salinexigens ZYF650T glucosylglycerol phosphorylase in glucosylglycerol catabolism
ComponentsSucrose phosphorylase
KeywordsSTRUCTURAL PROTEIN / Glucosylglycerol / phosphorylase-GH13_18 / structure-double displacement mechanism
Function / homology
Function and homology information


sucrose phosphorylase / sucrose phosphorylase activity / carbohydrate metabolic process
Similarity search - Function
Sucrose phosphorylase / Oligo-1,6-glucosidase, domain 2 / Alpha amylase, catalytic domain / Glycosyl hydrolase, family 13, catalytic domain / Alpha-amylase domain / Glycoside hydrolase superfamily
Similarity search - Domain/homology
alpha-D-glucopyranose / DI(HYDROXYETHYL)ETHER / Sucrose phosphorylase
Similarity search - Component
Biological speciesMarinobacter salinexigens (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.5 Å
AuthorsLu, D. / Ma, H.L.
Funding support China, 1items
OrganizationGrant numberCountry
National Natural Science Foundation of China (NSFC)32271358 China
CitationJournal: J Biol Chem / Year: 2025
Title: Structural basis of the bifunctionality of Marinobacter salinexigens ZYF650 glucosylglycerol phosphorylase in glucosylglycerol catabolism.
Authors: Di Lu / Keke Zhang / Chen Cheng / Danni Wu / Lei Yin / Quan Luo / Meiyun Shi / Honglei Ma / Xuefeng Lu /
Abstract: 2-O-α-Glucosylglycerol (GG) is a natural heteroside synthesized by many cyanobacteria and a few heterotrophic bacteria under salt stress conditions. Bacteria produce GG in response to stimuli and ...2-O-α-Glucosylglycerol (GG) is a natural heteroside synthesized by many cyanobacteria and a few heterotrophic bacteria under salt stress conditions. Bacteria produce GG in response to stimuli and degrade it once the stimulus diminishes. Heterotrophic bacteria utilize GG phosphorylase (GGP), a member of the GH13_18 family, via a two-step process consisting of phosphorolysis and hydrolysis for GG catabolism. However, the precise mechanism by which GGP degrades GG remains elusive. We determined the 3D structure of a recently identified GGP (MsGGP) of the deep-sea bacterium Marinobacter salinexigens ZYF650, in complex with glucose and glycerol, α-d-glucose-1-phosphate (αGlc1-P), and orthophosphate (inorganic phosphate) at resolutions of 2.5, 2.7, and 2.7 Å, respectively. Notably, the first αGlc1-P complex structure in the GH13_18 family, the complex of MsGGP and αGlc1-P, validates that GGP catalyzes GG decomposition through consecutive phosphorolysis and hydrolysis. In addition, the structure reveals the mechanism of high stereoselectivity on αGlc1-P. Glu231 and Asp190 were identified as the catalytic residues. Interestingly, these structures closely resemble each other, indicating minimal conformational changes upon binding end-product glucose and glycerol, or the intermediate αGlc1-P. The structures also indicate that the substrates may follow a specific trajectory and a precise order toward the active center in close proximity and in a geometrically favorable orientation for catalysis in a double displacement mechanism.
History
DepositionAug 6, 2024Deposition site: PDBJ / Processing site: PDBC
Revision 1.0Sep 10, 2025Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
B: Sucrose phosphorylase
D: Sucrose phosphorylase
C: Sucrose phosphorylase
A: Sucrose phosphorylase
E: Sucrose phosphorylase
F: Sucrose phosphorylase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)333,09740
Polymers329,4236
Non-polymers3,67434
Water22,1041227
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area25600 Å2
ΔGint-29 kcal/mol
Surface area94690 Å2
MethodPISA
Unit cell
Length a, b, c (Å)172.727, 115.065, 182.134
Angle α, β, γ (deg.)90.00, 100.56, 90.00
Int Tables number5
Space group name H-MC121
Components on special symmetry positions
IDModelComponents
11B-827-

HOH

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Components

#1: Protein
Sucrose phosphorylase / Glucosylglycerol phosphorylase


Mass: 54903.852 Da / Num. of mol.: 6
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Marinobacter salinexigens (bacteria) / Gene: gtfA, FWJ25_14990 / Production host: Escherichia coli (E. coli) / References: UniProt: A0A5B0VBK8, sucrose phosphorylase
#2: Chemical...
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 27 / Source method: obtained synthetically / Formula: C3H8O3 / Feature type: SUBJECT OF INVESTIGATION
#3: Sugar
ChemComp-GLC / alpha-D-glucopyranose / alpha-D-glucose / D-glucose / glucose


Type: D-saccharide, alpha linking / Mass: 180.156 Da / Num. of mol.: 6 / Source method: obtained synthetically / Formula: C6H12O6 / Feature type: SUBJECT OF INVESTIGATION
IdentifierTypeProgram
DGlcpaCONDENSED IUPAC CARBOHYDRATE SYMBOLGMML 1.0
a-D-glucopyranoseCOMMON NAMEGMML 1.0
a-D-GlcpIUPAC CARBOHYDRATE SYMBOLPDB-CARE 1.0
GlcSNFG CARBOHYDRATE SYMBOLGMML 1.0
#4: Chemical ChemComp-PEG / DI(HYDROXYETHYL)ETHER


Mass: 106.120 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C4H10O3
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1227 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY
Has protein modificationN

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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: 2.7 Å3/Da / Density % sol: 54.45 %
Crystal growTemperature: 289 K / Method: vapor diffusion / Details: 8% PEG 6000, 1.6 M Sodium chloride, 20% Glycerol

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: SSRF / Beamline: BL19U1 / Wavelength: 0.987 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Sep 17, 2023
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.987 Å / Relative weight: 1
ReflectionResolution: 2.29→54.8 Å / Num. obs: 157587 / % possible obs: 99.9 % / Redundancy: 6.8 % / Rmerge(I) obs: 0.128 / Rpim(I) all: 0.081 / Rrim(I) all: 0.152 / Net I/σ(I): 9.3
Reflection shellResolution: 2.29→2.35 Å / Num. unique obs: 11662 / Rpim(I) all: 0.822

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Processing

Software
NameVersionClassification
PHENIX(1.17.1_3660: ???)refinement
HKL-30007.21data scaling
HKL-30007.21data reduction
PHASER2.7.0phasing
PDB_EXTRACT3.27data extraction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 7XDQ
Resolution: 2.5→48.39 Å / SU ML: 0.29 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 21.19 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2115 5919 4.93 %
Rwork0.1743 --
obs0.1761 120051 99.06 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.5→48.39 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms23124 0 241 1227 24592
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00923919
X-RAY DIFFRACTIONf_angle_d1.26232461
X-RAY DIFFRACTIONf_dihedral_angle_d14.7823260
X-RAY DIFFRACTIONf_chiral_restr0.0683540
X-RAY DIFFRACTIONf_plane_restr0.0094217
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.5-2.530.29371960.24123613X-RAY DIFFRACTION94
2.53-2.560.2911750.24013667X-RAY DIFFRACTION97
2.56-2.590.31041960.24183728X-RAY DIFFRACTION97
2.59-2.620.25991750.22993809X-RAY DIFFRACTION99
2.62-2.660.26142120.22073803X-RAY DIFFRACTION100
2.66-2.690.25982070.2143761X-RAY DIFFRACTION100
2.69-2.730.27972270.21533845X-RAY DIFFRACTION100
2.73-2.770.26521990.2273766X-RAY DIFFRACTION100
2.77-2.820.27361950.22763848X-RAY DIFFRACTION100
2.82-2.860.30181940.22033808X-RAY DIFFRACTION100
2.86-2.910.29151930.22253838X-RAY DIFFRACTION100
2.91-2.960.25882060.21833881X-RAY DIFFRACTION100
2.96-3.020.26831960.21713769X-RAY DIFFRACTION100
3.02-3.080.25712280.20753803X-RAY DIFFRACTION100
3.08-3.150.22982060.20093850X-RAY DIFFRACTION100
3.15-3.220.22231940.19783806X-RAY DIFFRACTION100
3.22-3.30.23471810.19513843X-RAY DIFFRACTION100
3.3-3.390.20441720.18933866X-RAY DIFFRACTION100
3.39-3.490.21392110.17373828X-RAY DIFFRACTION100
3.49-3.610.21391970.17223797X-RAY DIFFRACTION100
3.61-3.730.22121890.16323857X-RAY DIFFRACTION100
3.73-3.880.1732120.15093789X-RAY DIFFRACTION100
3.88-4.060.17372160.13913841X-RAY DIFFRACTION100
4.06-4.270.17711880.13553835X-RAY DIFFRACTION100
4.27-4.540.15582040.12913839X-RAY DIFFRACTION99
4.54-4.890.18371840.1293832X-RAY DIFFRACTION99
4.89-5.380.15222000.13633807X-RAY DIFFRACTION99
5.38-6.160.19042180.15583816X-RAY DIFFRACTION99
6.16-7.760.17991730.1653840X-RAY DIFFRACTION98
7.76-48.390.1731750.1553747X-RAY DIFFRACTION94
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)
10.9655-0.30080.15983.39330.02961.221-0.0144-0.26240.01570.47010.0298-0.2740.02470.1454-0.03470.2428-0.0092-0.04580.3883-0.00340.2371-15.2779-57.455875.4342
21.6171-0.26090.71821.6436-0.18691.5847-0.1095-0.08970.35310.104-0.02-0.1581-0.31270.11510.09850.2805-0.0664-0.04230.2765-0.02740.3501-21.3917-39.135365.3356
32.2585-0.12690.51711.29980.18681.39150.0146-0.16590.05670.1376-0.03030.05860.00240.00650.01890.20460.01960.00480.2215-0.01070.2243-30.4559-65.389362.7038
43.7749-0.75710.04142.2883-0.13191.57980.10770.2028-0.5399-0.0169-0.0314-0.22060.16690.249-0.05830.33260.1027-0.0030.3089-0.05930.3788-17.4051-78.767350.3169
51.6067-0.53760.15291.21270.49761.8820.0470.5298-0.1175-0.1413-0.25680.2521-0.0553-0.520.13010.30450.0146-0.06510.5388-0.02170.2663-81.2433-76.468310.7396
61.37390.017-0.50390.69911.03142.36320.01030.18610.093-0.048-0.19350.2609-0.0138-0.58510.11480.24950.0059-0.05640.3891-0.02050.2871-87.1585-71.328525.5382
72.72280.02490.9230.87020.44031.07180.01250.16030.0239-0.10780.01690.09370.1640.0013-0.07140.29270.038-0.01610.24880.00480.2495-64.9586-79.858130.1044
85.1442.57741.46893.21151.27663.28540.00610.08440.23920.0314-0.05760.27290.0869-0.07490.06590.26960.08420.02250.2618-0.00030.2504-61.7254-72.42628.3518
92.00180.21820.3421.46910.78651.6983-0.03540.50310.3108-0.24360.1026-0.0832-0.12760.1923-0.06660.25280.02260.00510.39370.0560.2816-59.5984-72.173214.7568
102.26270.3549-0.85783.5718-0.32352.01180.07670.1952-0.61320.00820.1255-0.55720.55040.2807-0.19020.41210.144-0.04910.3269-0.07990.4522-49.6465-93.061623.0845
110.9111-0.4310.23891.4305-0.23371.3002-0.0172-0.5277-0.83790.223-0.0648-0.08590.5080.1308-0.00940.67780.0529-0.05010.41140.23610.6013-56.8353-101.837763.2821
121.07970.0235-0.47931.0748-0.24721.37080.0871-0.2114-0.50740.0326-0.0197-0.13260.41710.2027-0.02970.48860.0899-0.0790.24850.04350.4881-49.5478-97.272849.5356
131.7757-0.27780.12951.46320.11260.62150.002-0.3785-0.13540.18750.0062-0.08870.1997-0.0168-0.00750.3038-0.0145-0.01950.26830.06050.2434-65.2525-79.624960.1602
144.00971.7053-0.22546.0680.20432.93440.0729-0.0825-0.06930.1193-0.0640.6590.3911-0.4123-0.0870.2772-0.0444-0.01970.32390.0370.2832-86.0381-80.898652.0076
151.2686-0.0011-0.28810.79750.11851.6006-0.0277-0.76670.30020.4138-0.12880.25220.0473-0.45590.10650.41130.10310.07740.6939-0.1520.3957-82.0109-42.800679.6513
160.7455-0.9501-0.44391.78751.82863.4023-0.0611-0.4080.01230.2013-0.02020.2040.2414-0.4775-0.01670.3014-0.00490.0780.5292-0.06050.3698-89.266-53.900166.8489
170.87390.3731-0.05050.94610.010.4043-0.057-0.1730.27760.02720.01310.1605-0.1625-0.1886-0.01610.31820.0987-0.00130.3294-0.09050.3356-75.4477-38.666260.8099
184.7165-0.3626-1.57433.0210.16254.6964-0.0828-0.037-0.12010.0574-0.03830.0419-0.0611-0.25770.1130.21920.016-0.02280.2536-0.07570.2893-62.9275-44.255261.2334
191.81230.0372-0.09621.76760.65731.8033-0.0791-0.54080.13930.26990.0473-0.0991-0.03380.01070.03780.30410.076-0.02540.3848-0.08920.3142-60.2214-44.220474.6531
201.2686-0.0642-0.12766.04971.60112.2939-0.0332-0.36170.7278-0.15910.1355-0.1773-0.48870.0297-0.17080.42650.0152-0.05550.2976-0.14660.5983-53.4441-22.247366.1004
210.32460.3328-0.22190.9616-0.00971.3019-0.39260.60810.9222-0.2922-0.1398-0.2547-0.74290.0573-0.35470.7412-0.022-0.13120.36310.40120.8964-62.8927-14.427326.2498
220.3863-0.4640.79561.2221-0.40081.7208-0.23550.33310.9465-0.1087-0.0693-0.2601-0.50.37430.13050.5149-0.0578-0.07670.32580.17110.8829-49.515-17.198237.2787
230.71720.42410.1833.0777-0.17451.1664-0.04120.02180.41760.0011-0.0062-0.2079-0.2577-0.05750.0140.32870.0788-0.03310.2358-0.01010.4601-66.4475-31.634341.1964
242.4619-0.04950.20851.25240.32071.2277-0.00780.35890.1497-0.2407-0.040.0258-0.2886-0.2070.01960.34490.0802-0.01740.28470.03880.3201-75.9378-36.93729.373
251.23320.2227-0.25871.72120.01191.54770.03870.6544-0.0482-0.7995-0.1401-0.61180.09120.24970.02830.58490.12620.14320.8744-0.01080.3556-16.7431-60.398612.6599
261.31880.1471-0.45581.63-0.68320.8638-0.08370.4907-0.2668-0.37030.078-0.19590.22290.28130.03440.3960.09340.06930.6087-0.10830.3509-20.508-72.031223.302
271.2612-0.32540.35131.79290.29070.77420.04080.2596-0.0317-0.1056-0.12430.0311-0.08680.18940.07090.27620.01320.03080.42810.05010.2818-32.3532-51.271831.8323
281.4557-0.2173-0.10852.73670.25112.2396-0.02690.47990.3904-0.2424-0.1304-0.1537-0.34530.23680.1190.3242-0.01750.03210.49530.16620.4048-24.4008-38.023126.7125
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'B' and (resid 2 through 92 )
2X-RAY DIFFRACTION2chain 'B' and (resid 93 through 251 )
3X-RAY DIFFRACTION3chain 'B' and (resid 252 through 423 )
4X-RAY DIFFRACTION4chain 'B' and (resid 424 through 480 )
5X-RAY DIFFRACTION5chain 'D' and (resid 2 through 117 )
6X-RAY DIFFRACTION6chain 'D' and (resid 118 through 224 )
7X-RAY DIFFRACTION7chain 'D' and (resid 225 through 276 )
8X-RAY DIFFRACTION8chain 'D' and (resid 277 through 313 )
9X-RAY DIFFRACTION9chain 'D' and (resid 314 through 423 )
10X-RAY DIFFRACTION10chain 'D' and (resid 424 through 480 )
11X-RAY DIFFRACTION11chain 'C' and (resid 2 through 117 )
12X-RAY DIFFRACTION12chain 'C' and (resid 118 through 251 )
13X-RAY DIFFRACTION13chain 'C' and (resid 252 through 423 )
14X-RAY DIFFRACTION14chain 'C' and (resid 424 through 480 )
15X-RAY DIFFRACTION15chain 'A' and (resid 2 through 117 )
16X-RAY DIFFRACTION16chain 'A' and (resid 118 through 182 )
17X-RAY DIFFRACTION17chain 'A' and (resid 183 through 276 )
18X-RAY DIFFRACTION18chain 'A' and (resid 277 through 313 )
19X-RAY DIFFRACTION19chain 'A' and (resid 314 through 423 )
20X-RAY DIFFRACTION20chain 'A' and (resid 424 through 480 )
21X-RAY DIFFRACTION21chain 'E' and (resid 2 through 117 )
22X-RAY DIFFRACTION22chain 'E' and (resid 118 through 208 )
23X-RAY DIFFRACTION23chain 'E' and (resid 209 through 313 )
24X-RAY DIFFRACTION24chain 'E' and (resid 314 through 480 )
25X-RAY DIFFRACTION25chain 'F' and (resid 2 through 117 )
26X-RAY DIFFRACTION26chain 'F' and (resid 118 through 235 )
27X-RAY DIFFRACTION27chain 'F' and (resid 236 through 361 )
28X-RAY DIFFRACTION28chain 'F' and (resid 362 through 480 )

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