[English] 日本語
Yorodumi
- PDB-4hmj: Crystal structure of Staphylococcal nuclease variant Delta+PHS L3... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: PDB / ID: 4hmj
TitleCrystal structure of Staphylococcal nuclease variant Delta+PHS L36D at cryogenic temperature
ComponentsThermonuclease
KeywordsHYDROLASE / nuclease / hyperstable / pdTp / ionizable group
Function / homology
Function and homology information


micrococcal nuclease / endonuclease activity, active with either ribo- or deoxyribonucleic acids and producing 3'-phosphomonoesters / nucleic acid binding / extracellular region / membrane / metal ion binding
Similarity search - Function
OB fold (Dihydrolipoamide Acetyltransferase, E2P) - #90 / Thermonuclease family signature 1. / Thermonuclease active site / Thermonuclease family signature 2. / Staphylococcal nuclease (SNase-like), OB-fold / Staphylococcal nuclease homologue / Thermonuclease domain profile. / Staphylococcal nuclease homologues / SNase-like, OB-fold superfamily / OB fold (Dihydrolipoamide Acetyltransferase, E2P) ...OB fold (Dihydrolipoamide Acetyltransferase, E2P) - #90 / Thermonuclease family signature 1. / Thermonuclease active site / Thermonuclease family signature 2. / Staphylococcal nuclease (SNase-like), OB-fold / Staphylococcal nuclease homologue / Thermonuclease domain profile. / Staphylococcal nuclease homologues / SNase-like, OB-fold superfamily / OB fold (Dihydrolipoamide Acetyltransferase, E2P) / Beta Barrel / Mainly Beta
Similarity search - Domain/homology
THYMIDINE-3',5'-DIPHOSPHATE / Thermonuclease
Similarity search - Component
Biological speciesStaphylococcus aureus (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 1.35 Å
AuthorsRobinson, A.C. / Schlessman, J.L. / Heroux, A. / Garcia-Moreno E., B.
CitationJournal: To be Published
Title: Crystal structure of Staphylococcal nuclease variant Delta+PHS L36D at cryogenic temperature
Authors: Robinson, A.C. / Schlessman, J.L. / Heroux, A. / Garcia-Moreno E., B.
History
DepositionOct 18, 2012Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 31, 2012Provider: repository / Type: Initial release
Revision 1.1Sep 20, 2023Group: 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_initial_refinement_model / pdbx_struct_conn_angle / struct_conn / 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_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_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_ref_seq_dif.details / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

-
Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

-
Assembly

Deposited unit
A: Thermonuclease
hetero molecules


Theoretical massNumber of molelcules
Total (without water)16,5883
Polymers16,1451
Non-polymers4422
Water2,108117
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)31.168, 60.289, 38.342
Angle α, β, γ (deg.)90.000, 93.300, 90.000
Int Tables number4
Space group name H-MP1211

-
Components

#1: Protein Thermonuclease / TNase / Micrococcal nuclease / Staphylococcal nuclease


Mass: 16145.393 Da / Num. of mol.: 1 / Fragment: Nuclease A (UNP residues 83-231) / Mutation: L36D/G50F/V51N/P117G/H124L/S128A/Del44-49
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Staphylococcus aureus (bacteria) / Plasmid: pET24a+ / Production host: Escherichia coli (E. coli) / Strain (production host): BL21(DE3) / References: UniProt: P00644, micrococcal nuclease
#2: Chemical ChemComp-CA / CALCIUM ION


Mass: 40.078 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Ca
#3: Chemical ChemComp-THP / THYMIDINE-3',5'-DIPHOSPHATE


Type: DNA linking / Mass: 402.188 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C10H16N2O11P2
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 117 / Source method: isolated from a natural source / Formula: H2O

-
Experimental details

-
Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

-
Sample preparation

CrystalDensity Matthews: 2.23 Å3/Da / Density % sol: 44.78 %
Crystal growTemperature: 277 K / Method: vapor diffusion, hanging drop / pH: 8
Details: 20% MPD, 25 mM potassium phosphate, with calcium chloride and pdTp added to protein in a 3:2:1 ratio, pH 8, VAPOR DIFFUSION, HANGING DROP, temperature 277K

-
Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: NSLS / Beamline: X25 / Wavelength: 0.979 Å
DetectorType: DECTRIS PILATUS 6M / Detector: PIXEL / Date: Jul 17, 2012
Details: Meridionally-bent fused silica mirror with palladium and uncoated stripes vertically-focusing at 6.6:1 demagnification
RadiationMonochromator: double crystal Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.979 Å / Relative weight: 1
ReflectionResolution: 1.35→50 Å / Num. all: 30941 / Num. obs: 30941 / % possible obs: 99.8 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 6.6 % / Biso Wilson estimate: 23.2 Å2 / Rmerge(I) obs: 0.094 / Χ2: 2.966 / Net I/σ(I): 46.7
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsNum. unique allΧ2Diffraction-ID% possible all
1.35-1.376.50.30715661.0091100
1.37-1.46.50.27515201.051100
1.4-1.436.30.23715281.113199.9
1.43-1.456.70.21515611.241100
1.45-1.496.60.19815501.21100
1.49-1.526.20.17315151.3891100
1.52-1.566.80.16315481.4451100
1.56-1.66.70.1515371.6091100
1.6-1.656.60.13315711.8321100
1.65-1.76.40.12315201.9621100
1.7-1.766.80.11615452.0681100
1.76-1.836.60.10915912.4791100
1.83-1.926.70.10415272.9111100
1.92-2.026.90.115433.4841100
2.02-2.146.60.09515434.2371100
2.14-2.316.80.0915604.5351100
2.31-2.546.60.08715614.561199.9
2.54-2.916.90.08815345.087199.7
2.91-3.666.50.08615416.598198.2
3.66-506.50.09415809.112198.4

-
Phasing

PhasingMethod: molecular replacement
Phasing MRModel details: Phaser MODE: MR_AUTO
Highest resolutionLowest resolution
Rotation2.5 Å32.32 Å
Translation2.5 Å32.32 Å

-
Processing

Software
NameVersionClassificationNB
SCALEPACKdata scaling
PHASER2.2.1phasing
REFMACrefinement
PDB_EXTRACT3.11data extraction
CBASSdata collection
HKL-2000data reduction
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: PDB ENTRY 3BDC

3bdc


Resolution: 1.35→32.32 Å / Cor.coef. Fo:Fc: 0.967 / Cor.coef. Fo:Fc free: 0.959 / WRfactor Rfree: 0.2095 / WRfactor Rwork: 0.1911 / Occupancy max: 1 / Occupancy min: 0.4 / FOM work R set: 0.8892 / SU B: 1.441 / SU ML: 0.031 / SU R Cruickshank DPI: 0.0552 / SU Rfree: 0.0561 / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R: 0.055 / ESU R Free: 0.056 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : WITH TLS ADDED
RfactorNum. reflection% reflectionSelection details
Rfree0.1998 1552 5 %RANDOM
Rwork0.1797 ---
all0.1808 29368 --
obs0.1808 29368 99.3 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso max: 64.08 Å2 / Biso mean: 20.986 Å2 / Biso min: 10.36 Å2
Baniso -1Baniso -2Baniso -3
1--0.08 Å20 Å20.02 Å2
2---0.04 Å20 Å2
3---0.13 Å2
Refinement stepCycle: LAST / Resolution: 1.35→32.32 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms1033 0 26 117 1176
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.020.0221130
X-RAY DIFFRACTIONr_angle_refined_deg1.96821533
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.4555144
X-RAY DIFFRACTIONr_dihedral_angle_2_deg33.71125.09851
X-RAY DIFFRACTIONr_dihedral_angle_3_deg13.01415224
X-RAY DIFFRACTIONr_dihedral_angle_4_deg8.209155
X-RAY DIFFRACTIONr_chiral_restr0.1340.2165
X-RAY DIFFRACTIONr_gen_planes_refined0.0160.021831
X-RAY DIFFRACTIONr_mcbond_it1.3051.5663
X-RAY DIFFRACTIONr_mcangle_it2.02921071
X-RAY DIFFRACTIONr_scbond_it3.2673467
X-RAY DIFFRACTIONr_scangle_it4.6694.5454
LS refinement shellResolution: 1.35→1.385 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.227 101 -
Rwork0.241 2009 -
all-2110 -
obs-2110 93.03 %
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL11 (°2)L12 (°2)L13 (°2)L22 (°2)L23 (°2)L33 (°2)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T11 (Å2)T12 (Å2)T13 (Å2)T22 (Å2)T23 (Å2)T33 (Å2)Origin x (Å)Origin y (Å)Origin z (Å)
11.3214-0.4045-0.54242.5710.65930.378-0.0925-0.0421-0.17070.12240.09350.00140.06650.0802-0.00090.11720.0166-0.00610.0980.01220.085813.192-4.2413.372
21.4408-3.4712-0.87728.61471.92530.42450.04570.0684-0.0726-0.0897-0.0662-0.1496-0.0092-0.03270.02050.09560.01740.00370.0905-0.01820.115717.809-3.7990.048
31.9829-0.3141-0.55942.6983-0.17552.5814-0.0704-0.33850.19910.37490.0766-0.19-0.1230.273-0.00620.09310.0052-0.02950.1231-0.020.045113.50510.85710.584
43.26110.8804-0.08895.97852.32692.7969-0.1911-0.4828-0.3160.53810.1070.19760.26550.06890.08410.17880.04430.03060.130.07490.01368.767-0.29514.826
52.3474-0.0446-0.21673.8279-0.64672.26910.06020.2789-0.1212-0.1711-0.0780.0895-0.11570.06170.01780.1156-0.0034-0.00980.1131-0.01390.04769.097-0.3-4.269
62.59130.36450.24532.97750.24150.1948-0.01690.1378-0.1277-0.00420.0415-0.03050.030.0117-0.02460.11120.0034-0.00460.10930.00430.058912.1131.8720.061
71.6545-0.2601-0.28124.42120.33562.76-0.0925-0.19370.02910.2550.05690.24530.0646-0.06360.03560.0850.01710.02110.0960.01010.05393.3987.669.554
83.3295-0.4852-0.28122.38520.3670.25740.01540.26380.0615-0.2649-0.03230.0903-0.0727-0.02520.01690.08830.0004-0.01760.09340.02010.06273.65811.654-1.661
97.7662-0.59473.26794.4422-0.35557.7541-0.2326-0.50310.33620.47290.21760.2596-0.3677-0.29090.0150.12910.05360.04010.1025-0.04710.071-0.69415.84714.052
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A7 - 27
2X-RAY DIFFRACTION2A28 - 35
3X-RAY DIFFRACTION3A36 - 53
4X-RAY DIFFRACTION4A54 - 71
5X-RAY DIFFRACTION5A72 - 80
6X-RAY DIFFRACTION6A81 - 97
7X-RAY DIFFRACTION7A98 - 110
8X-RAY DIFFRACTION8A111 - 128
9X-RAY DIFFRACTION9A129 - 141

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242)：Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more