2F3J

The solution structure of the REF2-I mRNA export factor (residues 1-155).


SOLUTION NMR
NMR Experiment
ExperimentTypeSample ContentsSolventIonic StrengthpHPressureTemperature (K)Spectrometer
13D_13C-separated_NOESY13C,15N-REF2-I; 20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.0.126.3ambient303
23D_15N-separated_NOESY15N-REF2-I; 20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.0.126.3ambient303
32D NOESY15N-REF2-I; 20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O.0.126.3ambient303
4HSQC-IPAP15N,2H-REF2-I; 20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O, 5% liquid crystal media made of C12E5 and hexanol.20 mM Na phosphate buffer pH 6.3, 100 mM NaCl, 50 mM L-Arg, 50 mM L-Glu, 5 mM EDTA, 50 mM 2-mercaptoethanol, 10 mM DTT, 95% H2O, 5% D2O, 5% liquid crystal media made of C12E5 and hexanol.0.126.3ambient303
NMR Spectrometer Information
SpectrometerManufacturerModelField Strength
1BrukerDRX600
2VarianINOVA800
NMR Refinement
MethodDetailsSoftware
ARIA protocol (Nilges et al., 1997, J. Mol. Biol. 269, 408-422) used for structure calculation.Residual dipolar couplings were measured in 5% PEG liquid crystal media (C12E5 + hexane, Ruckert and Otting, 2000, J. Am. Chem. Soc. 122, 7793) and used as SANI restraints in ARIA.XwinNMR
NMR Ensemble Information
Conformer Selection Criteriastructures with the lowest energy
Conformers Calculated Total Number20
Conformers Submitted Total Number14
Representative Model1 (lowest energy)
Additional NMR Experimental Information
DetailsThe protein construct contains 14-residue N-terminal T7 tag (MASMTGGQQMGRDP), and C-terminal tag (LEHHHHHH). The T7 tag is unstructured and is omitted in the coordinate file, where residue numbering starts from residue 1 of REF2-I. Last three residues (LEH) in the coordinate file originate from the beginning of C-terminal tag. The N-terminal domain (1-74) of REF2-I is flexible and is largely unstructured, apart from the region 8-18 which forms a transient helix.
Computation: NMR Software
#ClassificationVersionSoftware NameAuthor
1collectionXwinNMR3.5Bruker
2processingNMRPipeDelaglio et al.
3data analysisNMRView5.1Johnson and Blevins
4structure solutionARIA1.1Nilges et al.
5structure solutionCNS1.0Brunger
6refinementCNS1.0Brunger