Experiment: 6F2X

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 900
2	2D 1H-15N HSQC	0.2 mM [U-13C; U-15N; U-2H] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
3	2D 1H-15N HSQC	0.3 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 800
4	3D HNHA	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
5	3D HNCA	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
6	3D HN(CO)CA	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
7	3D HNCACB	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
8	3D HN(COCA)CB	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
9	3D HNCO	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
10	3D HCC(CO)NH	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
11	3D CC(CO)NH	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
12	3D HCCH-TOCSY	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 600
13	3D 1H-15N NOESY	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 900
14	2D 1H-13C HSQC aliphatic	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
24	2D 1H-13C HSQC aromatic	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 900
17	3D 1H-13C NOESY aliphatic	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
18	3D 1H-13C NOESY aromatic	0.2 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 900
19	2D 1H-15N HETNOE	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 700
20	2D 1H-15N T1	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 700
21	2D 1H-15N T2	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 700
22	2D 1H-15N IPAP-HSQC	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 700
23	2D 1H-15N IPAP-HSQC	0.2 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 700
25	3D HNCACB	0.3 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 800
26	3D HN(CA)NNH	0.3 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 800
27	3D 1H-15N NOESY	0.3 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 800
28	3D HNCO	0.3 mM [U-13C; U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 800
29	3D HNHA	0.3 mM [U-15N] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	330 mM	2	ambient mbar	298	Bruker AVANCE 600
30	3D HNCACB	0.2 mM [U-13C; U-15N; U-2H] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
31	3D HN(COCA)CB	0.2 mM [U-13C; U-15N; U-2H] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
32	3D HNCO	0.2 mM [U-13C; U-15N; U-2H] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950
33	3D HN(CA)CO	0.2 mM [U-13C; U-15N; U-2H] PtkA, 50 mM HEPES, 300 mM sodium chloride, 10 mM DTT, 10 mM magnesium chloride	90% H2O/10% D2O	354 mM	7.5	ambient mbar	298	Bruker AVANCE 950

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600
2	Bruker	AVANCE	700
3	Bruker	AVANCE	800
4	Bruker	AVANCE	900
5	Bruker	AVANCE	950

NMR Refinement
Method	Details	Software
simulated annealing with torsion angle dynamics	structure determination	CYANA
cartesian angle dynamics	energy minimization	CNS
molecular dynamics	refinement in water	ARIA

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	collection	TopSpin	3.2	Bruker Biospin
2	processing	TopSpin	3.2	Bruker Biospin
3	chemical shift assignment	Sparky	3.114	Goddard and Kneller
4	peak picking	Sparky	3.114	Goddard and Kneller
5	data analysis	Sparky	3.114	Goddard and Kneller
6	structure calculation	CYANA	3.97	Guntert, Mumenthaler and Wuthrich
7	structure calculation	CNS	1.1	Brunger, Adams, Clore, Gros, Nilges and Read
8	refinement	ARIA	1.2 HJ development version	Linge, O'Donoghue and Nilges

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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6F2X

Structural characterization of the Mycobacterium tuberculosis Protein Tyrosine Kinase A (PtkA)