Experiment: 5T1O

SOLUTION NMR - SOLUTION SCATTERING

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HN(CO)CACB TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
2	3D HN(CA)CO TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
3	3D HNCACB TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
7	3D HNCO TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
8	3D HNCA TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
4	3D 1H-15N NOESY TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
9	3D HNCA TROSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
10	3D 1H-15N NOESY TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
11	3D HN(CO)CACB TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
12	3D HNCACB TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
13	3D HNCA TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
14	3D HNCACB	200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM na EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 600
15	ARTSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 900
16	ARTSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 900
17	ARTSY	200 uM [U-13C; U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA, 10 mg/mL Pf1 phage	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 900
18	ARTSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-13C; U-15N; U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA, 10 mg/mL Pf1 phage	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 900
19	2D 1H-15N HSQC TROSY	200 uM [U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
20	2D 1H-15N HSQC TROSY	200 uM [U-15N; U-2H] EIN-Ntr, 240 uM [U-2H] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
21	2D 1H-15N HSQC TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-2H; U-15N] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800
22	2D 1H-15N HSQC TROSY	240 uM [U-2H] EIN-Ntr, 200 uM [U-2H; U-15N] NPr, 10 mM TRIS, 100 mM sodium chloride, 0.5 mM EDTA	93% H2O/7% D2O	108 mM	7.5	1 atm	300	Bruker AVANCE 800

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600
2	Bruker	AVANCE	800
3	Bruker	AVANCE	900

NMR Refinement
Method	Details	Software
simulated annealing	DOCKING WAS FIRST CARRIED OUT BETWEEN THE TWO PROTEINS USING RIGID BODY SIMULATED ANNEALING (USING THE FREE FORM STRUCTURES - 5T1N AND 5T12). THE TOP TWENTY STRUCTURES WERE THEN TAKEN THROUGH FOR THE REFINEMENT STEP. TORSION ANGLE DYNAMICS REFINEMENT. IN THE REFINEMENT STEP ALL RESIDUES IN NPR (AS WELL AS THE PCS TAG) EXCEPT FOR RESIDUES 14-18 WERE RIGID. EIN WAS FLEXIBLE. NCS RESTRAINTS WERE USED TO GUIDE DOMAIN ORIENTATION OF EIN AND TO AID IN THE CONVERGENCE OF THE COMPLEX STRUCTURE.	X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	X-PLOR NIH	2.37.7	SCHWIETERS, KUSZEWSKI, TJANDRA AND CLORE
2	structure solution	ANALYSIS	2.4.2
3	structure solution	TopSpin	3.0
4	structure solution	Gaussian	9
5	structure solution	NMRPipe	8.2
6	structure solution	TALOS-N	4.12

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), 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.

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5T1O

Solution-state NMR and SAXS structural ensemble of NPr (1-85) in complex with EIN-Ntr (170-424)