2MIO

Solution NMR Structure of SH3 Domain 1 of Rho GTPase-activating Protein 10 from Homo sapiens, Northeast Structural Genomics Consortium (NESG) Target HR9129A

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
2	2D 1H-13C CT HSQC aliphatic	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
3	3D HNCO	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
4	3D CBCA(CO)NH	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
5	3D HNCACB	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
6	2D 1H-13C CT HSQC aromatic	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
7	3D simutaneous 13C-aromatic,13C-aliphatic,15N edited 1H-1H NOESY	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
8	2D 1H-13C HSQC aliphatic	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
9	2D 1H-13C HSQC aromatic	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
10	3D HBHA(CO)NH	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
11	3D(H)CCH-TOCSY ali	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
12	3D (H)CCH-COSY ali	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
13	GFT-43D (H)CCH-COSY aromatic	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
14	3D HN(CA)CO	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
15	gNfHSQC_His	0.72 mM 10% 13C U-100% 15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
16	2D 1H-15N HSQC	0.72 mM U-100% 13C-U-100%15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
17	2D 1H-13C ct-HSQC methyl(28ms)	0.72 mM U-100% 13C-U-100%15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
18	2D 1H-13C CT HSQC-methyl (42ms)	0.72 mM U-100% 13C-U-100%15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
19	2D 1H-13C CT HSQC-methyl (56ms)	0.72 mM U-100% 13C-U-100%15N labeled protein	90% H2O/10% D2O		6.5	ambient	298
20	2D J-modulation 1H-15N HSQC	0.72 mM U-100% 13C-U-100%15N labeled protein	90% H2O/10% D2O		6.5	ambient	298

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	600

NMR Refinement
Method	Details	Software
simulated annealing	Structure determination was performed by running CYANA and ASDP in parallel using NOE-based constraints and PHI and PSI dihedral angle constraints from TALOSN. Consensus peak assignments were selected and used in iterative refinement with CYANA. The 20 conformers out of 100 with the lowest target function were further refined by simulated annealing in explicit water bath using the program CNS with PARAM19 force field NMR.	CNS

NMR Ensemble Information
Conformer Selection Criteria	target function
Conformers Calculated Total Number	100
Conformers Submitted Total Number	20
Representative Model	1 (lowest energy)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
2	structure solution	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
3	geometry optimization	CNS		Brunger, Adams, Clore, Gros, Nilges and Read
4	refinement	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
5	geometry optimization	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
6	structure solution	CYANA	3.0	Guntert, Mumenthaler and Wuthrich
7	data analysis	AutoStructure	2.1	Huang, Tejero, Powers and Montelione
8	refinement	AutoStructure	2.1	Huang, Tejero, Powers and Montelione
9	data analysis	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
10	chemical shift assignment	AutoAssign	2.1	Zimmerman, Moseley, Kulikowski and Montelione
11	data analysis	XEASY		Bartels et al.
12	peak picking	XEASY		Bartels et al.
13	chemical shift assignment	XEASY		Bartels et al.
14	collection	VnmrJ		Varian
15	geometry optimization	TALOSN		Shen, Cornilescu, Delaglio and Bax
16	structure validation	PSVS		Bhattacharya, Montelione
17	processing	PSVS		Bhattacharya, Montelione
18	structure validation	PSVS		Guntert
19	processing	PSVS		Guntert
20	structure validation	PSVS		Bhattacharya, Montelione
21	processing	PSVS		Bhattacharya, Montelione
22	structure validation	PSVS		Guntert
23	processing	PSVS		Guntert
24	data analysis	CSI		David Wishart,Leigh Willard,Tim Jellard,Brian Sykes
25	data analysis	MOLMOL		Koradi, Billeter and Wuthrich

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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2MIO

Solution NMR Structure of SH3 Domain 1 of Rho GTPase-activating Protein 10 from Homo sapiens, Northeast Structural Genomics Consortium (NESG) Target HR9129A