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- PDB-2x4f: The Crystal Structure of the human myosin light chain kinase LOC3... -

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Basic information

Entry
Database: PDB / ID: 2x4f
TitleThe Crystal Structure of the human myosin light chain kinase LOC340156.
ComponentsMYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4
KeywordsTRANSFERASE / LUNG / KINASE / MYOSIN / BREAST CANCER
Function / homology
Function and homology information


myosin light chain kinase activity / non-specific serine/threonine protein kinase / ATP binding
Similarity search - Function
Transferase(Phosphotransferase) domain 1 / Transferase(Phosphotransferase); domain 1 / Phosphorylase Kinase; domain 1 / Phosphorylase Kinase; domain 1 / Serine/threonine-protein kinase, active site / Serine/Threonine protein kinases active-site signature. / Protein kinase domain / Serine/Threonine protein kinases, catalytic domain / Protein kinase, ATP binding site / Protein kinases ATP-binding region signature. ...Transferase(Phosphotransferase) domain 1 / Transferase(Phosphotransferase); domain 1 / Phosphorylase Kinase; domain 1 / Phosphorylase Kinase; domain 1 / Serine/threonine-protein kinase, active site / Serine/Threonine protein kinases active-site signature. / Protein kinase domain / Serine/Threonine protein kinases, catalytic domain / Protein kinase, ATP binding site / Protein kinases ATP-binding region signature. / Protein kinase domain profile. / Protein kinase domain / Protein kinase-like domain superfamily / 2-Layer Sandwich / Orthogonal Bundle / Mainly Alpha / Alpha Beta
Similarity search - Domain/homology
Chem-16X / Myosin light chain kinase family member 4
Similarity search - Component
Biological speciesHOMO SAPIENS (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.67 Å
AuthorsMuniz, J.R.C. / Mahajan, P. / Rellos, P. / Fedorov, O. / Shrestha, B. / Wang, J. / Elkins, J.M. / Daga, N. / Cocking, R. / Chaikuad, A. ...Muniz, J.R.C. / Mahajan, P. / Rellos, P. / Fedorov, O. / Shrestha, B. / Wang, J. / Elkins, J.M. / Daga, N. / Cocking, R. / Chaikuad, A. / Krojer, T. / Ugochukwu, E. / Yue, W. / von Delft, F. / Arrowsmith, C.H. / Edwards, A.M. / Weigelt, J. / Bountra, C. / Gileadi, O. / Knapp, S.
CitationJournal: To be Published
Title: The Crystal Structure of the Human Myosin Light Chain Kinase Loc340156
Authors: Muniz, J.R.C. / Mahajan, P. / Rellos, P. / Fedorov, O. / Shrestha, B. / Wang, J. / Elkins, J.M. / Daga, N. / Cocking, R. / Chaikuad, A. / Krojer, T. / Ugochukwu, E. / Yue, W. / von Delft, F. ...Authors: Muniz, J.R.C. / Mahajan, P. / Rellos, P. / Fedorov, O. / Shrestha, B. / Wang, J. / Elkins, J.M. / Daga, N. / Cocking, R. / Chaikuad, A. / Krojer, T. / Ugochukwu, E. / Yue, W. / von Delft, F. / Arrowsmith, C.H. / Edwards, A.M. / Weigelt, J. / Bountra, C. / Gileadi, O. / Knapp, S.
History
DepositionJan 29, 2010Deposition site: PDBE / Processing site: PDBE
Revision 1.0Feb 9, 2010Provider: repository / Type: Initial release
Revision 1.1Dec 5, 2012Group: Database references / Derived calculations ...Database references / Derived calculations / Other / Refinement description / Source and taxonomy / Structure summary / Version format compliance
Revision 1.2Jan 24, 2018Group: Database references / Category: citation_author / Item: _citation_author.name
Revision 1.3May 8, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Other
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_database_status / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _pdbx_database_status.status_code_sf / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: MYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4
B: MYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)86,1709
Polymers84,9932
Non-polymers1,1777
Water4,540252
1
A: MYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)43,2836
Polymers42,4961
Non-polymers7875
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: MYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4
hetero molecules


Theoretical massNumber of molelcules
Total (without water)42,8873
Polymers42,4961
Non-polymers3902
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)148.880, 70.650, 96.570
Angle α, β, γ (deg.)90.00, 118.96, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B

NCS domain segments:
Dom-IDComponent-IDEns-IDRefine codeAuth asym-IDAuth seq-ID
1111A65 - 176
2111B65 - 176
1214A177 - 186
2214B177 - 186
1314A187 - 237
2314B187 - 237
1411A238 - 355
2411B238 - 355

NCS oper: (Code: given
Matrix: (-0.5003, 0.2219, -0.8369), (-0.2325, -0.9655, -0.117), (-0.834, 0.136, 0.5347)
Vector: 99.3039, 104.7071, 61.0736)

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Components

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Protein , 1 types, 2 molecules AB

#1: Protein MYOSIN LIGHT CHAIN KINASE FAMILY MEMBER 4 / SERINE/THREONINE-PROTEIN KINASE SGK085 / SUGEN KINASE 85


Mass: 42496.262 Da / Num. of mol.: 2 / Fragment: LIGHT CHAIN, RESIDUES 40-388
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) HOMO SAPIENS (human) / Cell line (production host): SF9 / Production host: SPODOPTERA FRUGIPERDA (fall armyworm)
References: UniProt: Q86YV6, non-specific serine/threonine protein kinase

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Non-polymers , 5 types, 259 molecules

#2: Chemical ChemComp-16X / 4-(2-amino-4-methyl-1,3-thiazol-5-yl)-N-(3-dioxaziridin-3-ylphenyl)pyrimidin-2-amine


Mass: 328.349 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: C14H12N6O2S
#3: Chemical ChemComp-SO4 / SULFATE ION / Sulfate


Mass: 96.063 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: SO4
#4: Chemical ChemComp-1PE / PENTAETHYLENE GLYCOL / PEG400 / Polyethylene glycol


Mass: 238.278 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C10H22O6 / Comment: precipitant*YM
#5: Chemical ChemComp-EDO / 1,2-ETHANEDIOL / ETHYLENE GLYCOL / Ethylene glycol


Mass: 62.068 Da / Num. of mol.: 3 / Source method: obtained synthetically / Formula: C2H6O2
#6: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 252 / 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: 2.62 Å3/Da / Density % sol: 53.12 % / Description: NONE
Crystal growDetails: 2M AMMONIUM_SULFATE, 2.5% W/V PEG 400, PH 8.0 HEPES

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I03 / Wavelength: 0.9795
DetectorType: ADSC CCD / Detector: CCD / Date: Dec 9, 2009
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9795 Å / Relative weight: 1
ReflectionResolution: 2.67→65.13 Å / Num. obs: 25080 / % possible obs: 99.9 % / Observed criterion σ(I): 2 / Redundancy: 3.7 % / Rmerge(I) obs: 0.04 / Net I/σ(I): 8.4
Reflection shellResolution: 2.67→2.81 Å / Redundancy: 3.7 % / Rmerge(I) obs: 0.65 / Mean I/σ(I) obs: 2 / % possible all: 99.9

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Processing

Software
NameVersionClassification
REFMAC5.5.0089refinement
iMOSFLMdata reduction
SCALAdata scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 2.67→84.49 Å / Cor.coef. Fo:Fc: 0.935 / Cor.coef. Fo:Fc free: 0.904 / SU B: 22.377 / SU ML: 0.215 / Cross valid method: THROUGHOUT / ESU R: 0.559 / ESU R Free: 0.297 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. U VALUES RESIDUAL ONLY.
RfactorNum. reflection% reflectionSelection details
Rfree0.24105 1278 5.1 %RANDOM
Rwork0.19012 ---
obs0.19267 23801 99.78 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 20.744 Å2
Baniso -1Baniso -2Baniso -3
1--0.73 Å20 Å2-0.01 Å2
2--1.45 Å20 Å2
3----0.73 Å2
Refinement stepCycle: LAST / Resolution: 2.67→84.49 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4552 0 79 252 4883
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0150.0224742
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.5261.986417
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg5.2955577
X-RAY DIFFRACTIONr_dihedral_angle_2_deg39.97325.278216
X-RAY DIFFRACTIONr_dihedral_angle_3_deg17.06515822
X-RAY DIFFRACTIONr_dihedral_angle_4_deg21.9021518
X-RAY DIFFRACTIONr_chiral_restr0.0920.2714
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.0213556
X-RAY DIFFRACTIONr_gen_planes_other
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_it0.7491.52888
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it1.37224656
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it1.80631854
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it3.1484.51755
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Ens-ID: 1 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1A1719tight positional0.210.05
2B1719tight positional0.210.05
1A504medium positional0.250.5
2B504medium positional0.250.5
1A1719tight thermal0.20.5
2B1719tight thermal0.20.5
1A504medium thermal0.22
2B504medium thermal0.22
LS refinement shellResolution: 2.67→2.739 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.335 112 -
Rwork0.281 1729 -
obs--99.89 %
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)
17.46092.15570.49413.5739-2.12125.67490.07750.4447-0.0685-0.6628-0.39860.12070.20840.20790.32110.28410.0770.01160.4203-0.00970.16582.171645.103417.6701
210.37840.613-0.267215.97261.18018.42890.0285-0.474-1.35110.225-0.3477-1.10320.91650.79160.31910.33120.21230.10390.55780.22320.389894.977235.72623.6185
34.7017-0.55740.34958.5637-3.02226.88790.2582-0.32380.49980.1753-0.5371-0.8081-0.14971.10630.2790.4452-0.2359-0.03541.01020.0870.760695.947947.518232.9004
47.446-2.742-3.44155.69322.196817.60060.0769-0.48620.30990.2439-0.3752-0.7808-0.11141.06850.29840.0680.0377-0.00020.34430.17550.165594.963944.656522.1298
59.715-0.3506-0.29484.4605-2.09875.7207-0.1944-1.3253-1.70080.3934-0.05910.23590.81920.2430.25350.35210.06140.13040.3930.23490.326182.12434.662933.3606
66.6343-1.5158-0.8242.793-0.59144.86450.039-0.254-0.31790.0408-0.2011-0.19620.16160.67750.16210.13790.00040.05550.32580.08460.084383.481443.805525.3754
70.62491.45520.1875.36311.00621.75050.1039-0.05590.1113-0.0113-0.0104-0.2045-0.41270.5179-0.09350.1804-0.12170.07150.3750.00820.18674.719858.927631.7003
86.2590.0358-5.16777.1778-5.097110.6570.6065-0.62960.72380.4943-0.5549-0.7353-1.21671.7278-0.05150.2447-0.24440.03080.4693-0.10460.264882.070259.034330.3925
93.0338-0.1799-1.53660.2836-0.12075.2137-0.0117-0.4921-0.2973-0.01110.0492-0.02590.42490.5461-0.03750.09520.0209-0.01990.23550.040.075870.500343.664238.6252
102.0645-0.2659-0.21371.2128-0.96613.17290.1768-0.41430.10550.2973-0.1503-0.0722-0.47610.2396-0.02640.1477-0.09520.01920.24710.01320.029868.237554.207345.5479
113.76990.5042-0.95128.7881-3.88577.00990.1901-0.56160.77170.6299-0.10740.1017-1.20160.0129-0.08270.3664-0.03710.07450.2129-0.09360.189765.214766.948942.1226
123.82380.76631.83373.8558-1.52765.85980.01360.17520.5555-0.31910.14080.5232-0.4719-0.1043-0.15440.23740.01630.07030.15010.03330.185562.009162.56931.0602
135.3574-0.99131.01383.9318-0.92725.0030.02980.32060.18280.0165-0.116-0.0141-0.3228-0.03720.08610.1296-0.01040.07150.280.02010.060951.417452.6845-4.8185
141.57230.8427-0.77331.73831.21412.45990.14530.57520.1094-0.3428-0.42490.394-0.712-1.17040.27960.45270.2530.09930.90720.04980.408339.434854.00471.2925
155.8959-3.0292-3.82936.16283.08712.1363-0.03440.54820.1803-0.1048-0.06160.0878-0.0907-0.24290.0960.08080.00750.03690.27010.04150.12648.304653.3625-6.9057
1614.0728-14.6006-3.089123.8737-0.933115.1763-0.2283-0.55990.84941.04390.2350.1884-1.9439-0.5983-0.00670.56610.00270.0260.2132-0.12780.270248.96262.398711.0142
172.7376-0.89690.08182.4232-1.95815.3579-0.060.22670.13760.0365-0.0366-0.0345-0.2131-0.12190.09660.0737-0.01590.05210.2092-0.0270.071551.068249.53231.6469
1815.829313.56670.477823.627610.615210.2857-0.07860.67610.6906-0.7521-0.08890.9721-0.4197-0.61050.16760.1778-0.0856-0.01470.2317-0.09680.234534.620432.08065.4887
197.01726.73430.85068.95262.01971.21390.00020.3035-0.74760.2290.1146-0.53360.27840.1978-0.11480.16090.05640.00490.2563-0.03710.188152.109832.752710.8069
204.15610.69031.47412.83032.36129.1999-0.13960.1781-0.1375-0.035-0.06140.22790.0174-0.35150.20110.01450.00250.01720.10850.02260.05549.140743.976912.1466
211.50410.16350.18372.07241.39093.17730.0776-0.16740.03080.1506-0.25710.20220.0206-0.60820.17950.0626-0.0250.03110.2857-0.01040.033439.606740.123923.2278
2232.7930.82723.349134.01215.988833.4133-0.0782-0.161-0.11810.4501-0.29681.60730.5244-1.82290.3750.2775-0.24790.11010.3465-0.0190.335632.810123.143319.8946
235.21520.6843-0.49555.26121.77084.92850.0118-0.3005-0.59130.47460.0434-0.29780.65990.1201-0.05520.18980.0042-0.01950.19360.10940.114149.256128.622722.6178
240.65771.68020.31254.30340.7950.44840.13030.0111-0.24190.29910.0488-0.54670.43490.1545-0.17910.7228-0.0013-0.23990.22910.05160.567252.003118.469612.4628
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A67 - 81
2X-RAY DIFFRACTION2A82 - 92
3X-RAY DIFFRACTION3A93 - 103
4X-RAY DIFFRACTION4A104 - 117
5X-RAY DIFFRACTION5A118 - 136
6X-RAY DIFFRACTION6A137 - 173
7X-RAY DIFFRACTION7A174 - 213
8X-RAY DIFFRACTION8A214 - 229
9X-RAY DIFFRACTION9A230 - 255
10X-RAY DIFFRACTION10A256 - 304
11X-RAY DIFFRACTION11A305 - 330
12X-RAY DIFFRACTION12A331 - 350
13X-RAY DIFFRACTION13B67 - 99
14X-RAY DIFFRACTION14B100 - 106
15X-RAY DIFFRACTION15B107 - 124
16X-RAY DIFFRACTION16B125 - 135
17X-RAY DIFFRACTION17B136 - 174
18X-RAY DIFFRACTION18B175 - 183
19X-RAY DIFFRACTION19B184 - 207
20X-RAY DIFFRACTION20B208 - 246
21X-RAY DIFFRACTION21B247 - 307
22X-RAY DIFFRACTION22B308 - 313
23X-RAY DIFFRACTION23B314 - 345
24X-RAY DIFFRACTION24B346 - 351

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