2JWZ

Mutations in the hydrophobic core of ubiquitin differentially affect its recognition by receptor proteins

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
2	2D 1H-1H TOCSY	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
3	2D 1H-1H NOESY	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
4	3D 15N-SEPARATED TOCSY	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
5	2D 1H-15N IPAP	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
6	HMQC-J	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
7	15N R1 AND R2	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296
8	15N{1H} NOE	2 mM [U-100% 15N] L69S UBIQUITIN, 2 mM L69S UBIQUITIN	93% H2O/7% D2O	20 mM	6.8	AMBIENT	296

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600

NMR Refinement
Method	Details	Software
simulated annealing		ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	75
Conformers Submitted Total Number	10
Representative Model	1 (lowest energy)

Additional NMR Experimental Information
Details	RESIDUAL DIPOLAR COUPLINGS WERE MEASURED IN POLYETHYLENE GLYCOL BASED ALIGNMENT MEDIUM

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	ARIA	2.0	Linge, O'Donoghue and Nilges
2	refinement	ARIA	2.0	Linge, O'Donoghue and Nilges
3	processing	CNS		Brunger, Adams, Clore, Gros, Nilges and Read

RCSB PDB Core Operations are funded by the U.S. National Science Foundation (DBI-2321666), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.