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- PDB-6x5h: Human Alpha-1,6-fucosyltransferase (FUT8) bound to GDP -

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Basic information

Entry
Database: PDB / ID: 6x5h
TitleHuman Alpha-1,6-fucosyltransferase (FUT8) bound to GDP
ComponentsAlpha-(1,6)-fucosyltransferase
KeywordsTRANSFERASE / Glycosyl transferase / Fucosyl transferase / GT-B fold / Inverting
Function / homology
Function and homology information


glycoprotein 6-alpha-L-fucosyltransferase / glycoprotein 6-alpha-L-fucosyltransferase activity / receptor metabolic process / GDP-L-fucose metabolic process / alpha-(1->6)-fucosyltransferase activity / N-glycan fucosylation / Reactions specific to the complex N-glycan synthesis pathway / L-fucose catabolic process / oligosaccharide biosynthetic process / regulation of cellular response to oxidative stress ...glycoprotein 6-alpha-L-fucosyltransferase / glycoprotein 6-alpha-L-fucosyltransferase activity / receptor metabolic process / GDP-L-fucose metabolic process / alpha-(1->6)-fucosyltransferase activity / N-glycan fucosylation / Reactions specific to the complex N-glycan synthesis pathway / L-fucose catabolic process / oligosaccharide biosynthetic process / regulation of cellular response to oxidative stress / N-glycan processing / respiratory gaseous exchange by respiratory system / protein N-linked glycosylation via asparagine / fibroblast migration / protein N-linked glycosylation / Golgi cisterna membrane / transforming growth factor beta receptor signaling pathway / integrin-mediated signaling pathway / SH3 domain binding / regulation of gene expression / Maturation of spike protein / in utero embryonic development / viral protein processing / Golgi membrane / Golgi apparatus / extracellular exosome / membrane
Similarity search - Function
Alpha-(1,6)-fucosyltransferase / Alpha-(1,6)-fucosyltransferase, SH3 domain / Alpha-(1,6)-fucosyltransferase, N- and catalytic domain / Alpha-(1,6)-fucosyltransferase N- and catalytic domains / Glycosyltransferase family 23 (GT23) domain / Glycosyltransferase family 23 (GT23) domain profile. / Variant SH3 domain / Src homology 3 domains / SH3-like domain superfamily / Src homology 3 (SH3) domain profile. / SH3 domain
Similarity search - Domain/homology
GUANOSINE-5'-DIPHOSPHATE / Alpha-(1,6)-fucosyltransferase
Similarity search - Component
Biological speciesHomo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.25 Å
AuthorsKadirvelraj, R. / Wood, Z.A.
Funding support United States, 1items
OrganizationGrant numberCountry
National Institutes of Health/National Institute on Alcohol Abuse and Alcoholism (NIH/NIAAA)R01GM130915, P41GM103390 and P01GM107012 United States
CitationJournal: J.Biol.Chem. / Year: 2020
Title: Characterizing human alpha-1,6-fucosyltransferase (FUT8) substrate specificity and structural similarities with related fucosyltransferases.
Authors: Boruah, B.M. / Kadirvelraj, R. / Liu, L. / Ramiah, A. / Li, C. / Zong, G. / Bosman, G.P. / Yang, J.Y. / Wang, L.X. / Boons, G.J. / Wood, Z.A. / Moremen, K.W.
History
DepositionMay 26, 2020Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 7, 2020Provider: repository / Type: Initial release
Revision 1.1Oct 21, 2020Group: Database references / Category: citation / citation_author
Item: _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title
Revision 1.2Dec 23, 2020Group: Database references / Category: citation / citation_author
Item: _citation.journal_volume / _citation.page_first ..._citation.journal_volume / _citation.page_first / _citation.page_last / _citation_author.identifier_ORCID
Revision 1.3Oct 18, 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
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: Alpha-(1,6)-fucosyltransferase
B: Alpha-(1,6)-fucosyltransferase
C: Alpha-(1,6)-fucosyltransferase
D: Alpha-(1,6)-fucosyltransferase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)249,70021
Polymers247,3814
Non-polymers2,31917
Water23,4011299
1
A: Alpha-(1,6)-fucosyltransferase
hetero molecules

D: Alpha-(1,6)-fucosyltransferase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)124,72212
Polymers123,6902
Non-polymers1,03210
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
crystal symmetry operation5_565y,-x+y+1,z+1/61
Buried area8470 Å2
ΔGint-21 kcal/mol
Surface area39690 Å2
MethodPISA
2
B: Alpha-(1,6)-fucosyltransferase
C: Alpha-(1,6)-fucosyltransferase
hetero molecules


Theoretical massNumber of molelcules
Total (without water)124,9779
Polymers123,6902
Non-polymers1,2877
Water362
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area8830 Å2
ΔGint-39 kcal/mol
Surface area38730 Å2
MethodPISA
Unit cell
Length a, b, c (Å)173.490, 173.490, 207.420
Angle α, β, γ (deg.)90.00, 90.00, 120.00
Int Tables number170
Space group name H-MP65

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Components

#1: Protein
Alpha-(1,6)-fucosyltransferase / Alpha1-6FucT / Fucosyltransferase 8 / GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha1 / 6- ...Alpha1-6FucT / Fucosyltransferase 8 / GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha1 / 6-fucosyltransferase / GDP-fucose--glycoprotein fucosyltransferase / Glycoprotein 6-alpha-L-fucosyltransferase


Mass: 61845.219 Da / Num. of mol.: 4
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: FUT8 / Cell (production host): HEK293 / Production host: Homo sapiens (human)
References: UniProt: Q9BYC5, glycoprotein 6-alpha-L-fucosyltransferase
#2: Chemical ChemComp-GDP / GUANOSINE-5'-DIPHOSPHATE


Type: RNA linking / Mass: 443.201 Da / Num. of mol.: 3 / Source method: isolated from a natural source / Formula: C10H15N5O11P2 / Feature type: SUBJECT OF INVESTIGATION / Comment: GDP, energy-carrying molecule*YM
#3: Chemical
ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL


Mass: 62.068 Da / Num. of mol.: 10 / Source method: isolated from a natural source / Formula: C2H6O2
#4: Chemical
ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 4 / Source method: isolated from a natural source / Formula: C3H8O3
#5: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 1299 / Source method: isolated from a natural source / Formula: H2O
Has ligand of interestY

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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: 3.75 Å3/Da / Density % sol: 67 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 7.5
Details: 1.6 M Ammonium sulfate, 0.6 M L-proline, 100 mM Hepes pH 7.5
Temp details: Room temperature

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 22-BM / Wavelength: 1 Å
DetectorType: MARRESEARCH / Detector: CCD / Date: Jun 21, 2017
RadiationMonochromator: Si (111) Rosenbaum-Rock double crystal / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 2.25→86.75 Å / Num. obs: 165616 / % possible obs: 99.2 % / Redundancy: 5.1 % / Biso Wilson estimate: 31.7 Å2 / CC1/2: 0.995 / Rrim(I) all: 0.158 / Net I/σ(I): 10.44
Reflection shellResolution: 2.25→2.31 Å / Redundancy: 4.1 % / Num. unique obs: 11970 / CC1/2: 0.556 / Rrim(I) all: 1.1 / % possible all: 97.1

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Processing

Software
NameVersionClassification
PHENIX1.12_2829refinement
XDSdata reduction
XSCALEdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2DE0
Resolution: 2.25→86.745 Å / SU ML: 0.25 / Cross valid method: FREE R-VALUE / σ(F): 1.36 / Phase error: 20.12 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.202 8312 5.02 %Random selection
Rwork0.1706 ---
obs0.1721 165569 99.17 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.25→86.745 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms14933 0 148 1301 16382
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00815554
X-RAY DIFFRACTIONf_angle_d0.91821100
X-RAY DIFFRACTIONf_dihedral_angle_d10.9459293
X-RAY DIFFRACTIONf_chiral_restr0.0532233
X-RAY DIFFRACTIONf_plane_restr0.0062701
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.2503-2.27580.32412610.28554929X-RAY DIFFRACTION94
2.2758-2.30260.29642730.2415235X-RAY DIFFRACTION100
2.3026-2.33070.2762840.22115314X-RAY DIFFRACTION100
2.3307-2.36020.27112690.22685241X-RAY DIFFRACTION100
2.3602-2.39130.25842850.22065260X-RAY DIFFRACTION100
2.3913-2.4240.25082820.20745287X-RAY DIFFRACTION100
2.424-2.45870.26432730.21515267X-RAY DIFFRACTION100
2.4587-2.49540.26552810.20795267X-RAY DIFFRACTION100
2.4954-2.53440.23892830.19775226X-RAY DIFFRACTION100
2.5344-2.57590.22842770.19585274X-RAY DIFFRACTION100
2.5759-2.62030.24392830.19665241X-RAY DIFFRACTION100
2.6203-2.6680.26472800.20195327X-RAY DIFFRACTION100
2.668-2.71930.22142670.18785216X-RAY DIFFRACTION100
2.7193-2.77480.21322780.18245276X-RAY DIFFRACTION100
2.7748-2.83510.23372780.18025265X-RAY DIFFRACTION100
2.8351-2.90110.22792810.17985263X-RAY DIFFRACTION100
2.9011-2.97370.212690.17515258X-RAY DIFFRACTION100
2.9737-3.05410.22452750.17835253X-RAY DIFFRACTION99
3.0541-3.14390.21162770.18135279X-RAY DIFFRACTION99
3.1439-3.24540.23212800.17945226X-RAY DIFFRACTION99
3.2454-3.36140.20082760.16825267X-RAY DIFFRACTION99
3.3614-3.4960.1952780.16425227X-RAY DIFFRACTION99
3.496-3.65510.18932760.14755282X-RAY DIFFRACTION99
3.6551-3.84780.17342750.14465250X-RAY DIFFRACTION99
3.8478-4.08890.15822840.1375223X-RAY DIFFRACTION99
4.0889-4.40460.16042820.12885190X-RAY DIFFRACTION99
4.4046-4.84780.142760.12415249X-RAY DIFFRACTION98
4.8478-5.54920.16232750.15065228X-RAY DIFFRACTION99
5.5492-6.99090.19212830.17435211X-RAY DIFFRACTION98
6.9909-86.7450.17342710.17655226X-RAY DIFFRACTION97
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)
11.27980.2451-0.34340.479-0.03750.6172-0.0883-0.02230.0409-0.00710.04110.0351-0.01660.0450.04290.24660.0046-0.02030.2342-0.04260.2278137.175716.895-10.0262
22.85380.6892-1.52021.3002-0.19872.1679-0.09930.1578-0.1339-0.03290.02940.04510.0218-0.10540.07920.2276-0.0439-0.00830.2269-0.0760.1951129.30012.3021-21.2761
30.65550.0203-0.14230.62210.10080.6097-0.063-0.0915-0.04450.0630.02920.01510.06080.03680.03090.24070.01290.00710.2634-0.02890.2617135.8296.537-7.8094
45.3018-1.0645-1.72442.181-0.0412.8117-0.07240.03240.0837-0.1768-0.0607-0.36410.00050.27960.13320.3289-0.0679-0.06470.25160.07590.3734197.204358.6279-57.8703
58.14463.9785-3.02694.531-3.34023.11810.35510.00080.82630.5042-0.08820.781-0.381-0.1545-0.19830.22790.046-0.06390.2612-0.0540.2477164.120249.7361-43.9047
62.25030.31220.54380.7690.06691.31040.0091-0.156-0.00040.0024-0.05030.04740.105-0.10980.04820.2729-0.0434-0.02340.26270.01040.2497163.909126.8345-37.5217
71.41140.65090.1321.69240.34551.1202-0.04040.00340.0251-0.1538-0.01070.08480.1461-0.03590.04780.2243-0.0231-0.06590.269-0.00670.2183162.923631.1779-48.0035
83.52830.6949-0.43042.1629-1.55361.41260.1059-0.1708-0.0194-0.0011-0.2105-0.086-0.13280.0540.0960.2108-0.0461-0.08830.25110.04530.1808185.564836.4642-33.2702
91.5580.9284-0.48982.088-1.49831.8951-0.0198-0.1297-0.01580.0466-0.0805-0.0881-0.02460.10020.11190.202-0.012-0.04920.2520.01610.2057191.531938.9827-34.3255
101.50180.1377-0.20350.9172-0.2570.8974-0.06870.1889-0.0148-0.23190.0133-0.04830.08830.03560.05210.3085-0.024-0.03380.23390.02390.2394180.456238.4319-57.4282
117.7494-3.4222-0.49424.06970.07222.0093-0.05850.39420.0742-0.3245-0.0892-0.32710.08940.02460.13940.3088-0.0895-0.02030.20950.04290.2354192.371350.6734-64.0417
121.4174-0.99920.13783.13990.1210.70920.02570.44580.5657-0.4967-0.1466-0.2658-0.2686-0.1131-0.01370.5690.0225-0.10480.44660.31080.646185.773892.1408-72.1337
131.2683-0.40730.38452.5747-0.93922.59950.03970.73670.2543-0.8905-0.0412-0.2612-0.0621-0.00430.05330.74350.03010.04520.63730.18320.3836193.611970.5465-84.7522
141.1628-0.64480.35462.0048-0.76711.3461-0.04130.08780.2793-0.1314-0.047-0.1669-0.20910.09210.09820.3583-0.0467-0.04520.28020.13060.4748196.5776.7435-59.8017
151.83180.704-0.60531.2148-0.0781.0341-0.0062-0.2303-0.20790.1584-0.01270.21220.0402-0.15-0.0260.2748-0.02150.08290.31940.06020.33210.601198.3555-25.3097
163.7834-0.21471.84121.10961.58633.6273-0.2321-0.3871-0.53360.39760.41320.33480.6232-0.2523-0.09310.4855-0.08020.29580.78720.14450.7933185.251482.9639-12.0426
172.4380.96190.07221.79170.2211.0456-0.20340.1657-0.5525-0.0420.14310.31220.3627-0.5591-0.07880.3356-0.17240.1330.55160.05120.6934190.292884.1039-27.3168
183.43511.0727-1.30111.5354-0.57493.00610.1569-0.7231-0.24070.5308-0.2360.11410.04690.21970.09160.4424-0.02010.09110.54650.08220.3347214.133995.267-6.1256
191.8548-0.4195-0.8731.09970.61122.5048-0.116-0.2867-0.17290.20810.0530.15670.3006-0.08370.09320.2748-0.03850.08180.29740.08350.4021211.02487.4242-23.3422
203.8413-0.0671-0.79240.0454-0.32121.17130.16220.0574-0.1972-0.052-0.00430.23870.1086-0.3581-0.12920.288-0.08310.0660.39260.01610.5168203.296289.2455-35.849
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 108 through 339 )
2X-RAY DIFFRACTION2chain 'A' and (resid 340 through 412 )
3X-RAY DIFFRACTION3chain 'A' and (resid 413 through 574 )
4X-RAY DIFFRACTION4chain 'B' and (resid 108 through 172 )
5X-RAY DIFFRACTION5chain 'B' and (resid 173 through 199 )
6X-RAY DIFFRACTION6chain 'B' and (resid 200 through 267 )
7X-RAY DIFFRACTION7chain 'B' and (resid 268 through 339 )
8X-RAY DIFFRACTION8chain 'B' and (resid 340 through 378 )
9X-RAY DIFFRACTION9chain 'B' and (resid 379 through 458 )
10X-RAY DIFFRACTION10chain 'B' and (resid 459 through 574 )
11X-RAY DIFFRACTION11chain 'C' and (resid 108 through 172 )
12X-RAY DIFFRACTION12chain 'C' and (resid 173 through 378 )
13X-RAY DIFFRACTION13chain 'C' and (resid 379 through 469 )
14X-RAY DIFFRACTION14chain 'C' and (resid 470 through 574 )
15X-RAY DIFFRACTION15chain 'D' and (resid 108 through 237 )
16X-RAY DIFFRACTION16chain 'D' and (resid 238 through 267 )
17X-RAY DIFFRACTION17chain 'D' and (resid 268 through 339 )
18X-RAY DIFFRACTION18chain 'D' and (resid 340 through 427 )
19X-RAY DIFFRACTION19chain 'D' and (resid 428 through 548 )
20X-RAY DIFFRACTION20chain 'D' and (resid 549 through 574 )

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