2KYM

Solution structure of the Bem1p SH3-CI domain from L.elongisporus in complex with Ste20p peptide

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
2	3D HNCO	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
3	3D HN(CO)CA	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
4	3D HNCACB	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
5	3D CBCA(CO)NH	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
6	3D C(CO)NH	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
7	3D HNCA	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	95% H2O/5% D2O	100	6.8	ambient	293
8	2D 1H-13C HSQC	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	100% D2O	100	6.8	ambient	293
9	3D HCCH-TOCSY	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	100% D2O	100	6.8	ambient	293
10	3D HCCH-COSY	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	100% D2O	100	6.8	ambient	293
11	3D 1H-15N NOESY	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	100% D2O	100	6.8	ambient	293
12	3D 1H-13C NOESY	0.7 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 1.5 mM Ste20 peptide, 0.5 mM PMSF, 0.5 mM EDTA, 0.05 % sodium azide, 50 mM HEPES, 100 mM sodium chloride, 1 mM DTT	100% D2O	100	6.8	ambient	293
13	15N C13 filtered 2D 1H-1H TOCSY	0.5 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 0.5 mM Ste20 peptide, 100 mM sodium chloride, 50 mM sodium phosphate, 0.05 % sodium azide	95% H2O/5% D2O	100	6.8	ambient	293
14	15N C13 filtered 2D 1H-1H NOESY	0.5 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 0.5 mM Ste20 peptide, 100 mM sodium chloride, 50 mM sodium phosphate, 0.05 % sodium azide	95% H2O/5% D2O	100	6.8	ambient	293
15	15N C13 filtered 2D 1H-1H COSY	0.5 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 0.5 mM Ste20 peptide, 100 mM sodium chloride, 50 mM sodium phosphate, 0.05 % sodium azide	100% D2O	100	6.8	ambient	293
16	13C half-filtered 1H-13C NOESY	0.5 mM [U-99% 13C; U-99% 15N] Bem1 SH3-CI protein, 0.5 mM Ste20 peptide, 100 mM sodium chloride, 50 mM sodium phosphate, 0.05 % sodium azide	100% D2O	100	6.8	ambient	293

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	800

NMR Refinement
Method	Details	Software
simulated annealing		Sparky

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	chemical shift assignment	Sparky		Goddard
2	peak picking	Sparky		Goddard
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	structure solution	CYANA	2.1	Guntert, Mumenthaler and Wuthrich
5	refinement	CYANA	2.1	Guntert, Mumenthaler 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.