2M68

NMR solution structure ensemble of 3-4D mutant domain 11 IGF2R in complex with IGF2 (domain 11 structure only)

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
2	2D 1H-13C HSQC	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
3	3D CBCA(CO)NH	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
4	3D C(CO)NH	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
5	3D HNCO	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
6	3D HNCA	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
7	3D HN(CO)CA	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
8	3D H(CCO)NH	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
9	3D HCCH-TOCSY	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
10	3D 1H-15N NOESY	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
11	3D 1H-15N TOCSY	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
12	3D 1H-13C NOESY aliphatic	STRUCTURAL STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	310.15
13	Heteronuclear NOE ratio	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15
14	Heteronuclear NOE ratio	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15
15	T1 experiments	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15
16	T1 experiments	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15
17	T2 experiments	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15
18	T2 experiments	DYNAMICS STUDIES: Lyophilised IGF2 was added to solution state domain 11 at pH 4 in a 1:1 ratio.	93% H2O/7% D2O	0.005	4	1 atm	298.15

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	VNMRS	600
2	Varian	VNMRS	900

NMR Refinement
Method	Details	Software
DGSA-distance geometry simulated annealing		ARIA

NMR Ensemble Information
Conformer Selection Criteria	structures with the least restraint violations
Conformers Calculated Total Number	200
Conformers Submitted Total Number	20
Representative Model	1 (fewest violations)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	processing	ARIA	2.2	Linge, O'Donoghue and Nilges
2	refinement	ARIA	2.2	Linge, O'Donoghue and Nilges
3	structure solution	ARIA	2.2	Linge, O'Donoghue and Nilges
4	refinement	CING	iCing	Doreleijers
5	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
6	processing	NMRDraw		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
7	data analysis	relax	1.3.9	Palmer
8	processing	relax	1.3.9	Palmer
9	data analysis	Sparky	2.6	Goddard
10	collection	VnmrJ	0.4	Varian
11	chemical shift assignment	CCPN_Analysis	2.1	CCPN
12	data analysis	CCPN_Analysis	2.1	CCPN
13	peak picking	CCPN_Analysis	2.1	CCPN
14	geometry optimization	TALOS	TALOS+	Cornilescu, Delaglio and Bax

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.