2KBN

Solution NMR structure of the OB domain (residues 67-166) of MM0293 from Methanosarcina mazei. Northeast Structural Genomics Consortium target MaR214a.


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
13D HNCO1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
23D HNCA1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
33D HNCACB1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
43D CBCA(CO)NH1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
53D HCCH-TOCSY1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
63D H(CCO)NH1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
73D C(CO)NH1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
83D 1H-15N NOESY1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
93D 1H-13C NOESY1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
102D 1H-15N HSQC1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
112D 1H-13C HSQC1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
122D 1H-13C HSQC1.2 mM [U-5% 13C; U-99% 15N] OB domain-7, 20 mM MES-8, 100 mM sodium chloride-9, 10 mM DTT-10, 5 mM calcium chloride-11, .02 % sodium azide-1295% H2O/5% D2O.16.5ambient293
134D 1H-13C NOESY1.2 mM [U-100% 13C; U-100% 15N] OB domain-1, 20 mM MES-2, 100 mM sodium chloride-3, 10 mM DTT-4, 5 mM calcium chloride-5, .02 % sodium azide-695% H2O/5% D2O.16.5ambient293
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerAVANCE850
2VarianINOVA600
NMR Refinement
MethodDetailsSoftware
simulated annealingautostructure and cyana were used to automatically assign NOE crosspeaks. xplor-NIH and CNS were used to do hydrogen bond refinement and refinement in the presence of water.TopSpin
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number25
Conformers Submitted Total Number20
Representative Model1 (lowest energy)
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1collectionTopSpin2.1Bruker Biospin
2data analysisSparky3.113Goddard
3chemical shift assignmentAutoAssign2.3.0Zimmerman, Moseley, Kulikowski and Montelione
4structure solutionAutoStructure2.2.1Huang, Tejero, Powers and Montelione
5data analysisCYANA2.1Guntert, Mumenthaler and Wuthrich
6data analysisPSVS1.3Bhattacharya and Montelione
7refinementCNS1.2Brunger, Adams, Clore, Gros, Nilges and Read
8refinementX-PLOR NIH2.2.1Schwieters, Kuszewski, Tjandra and Clore