ISDA: 2M0H

SOLUTION NMR

NMR Experiment

Experiment	Type	Sample Contents	Ionic Strength	Solvent	pH	Pressure	Temprature (K)
1	2D 1H-1H TOCSY	1.2 mM protein, 20 mM TCEP, methanol	20	methanol	4.0	ambient	?
2	2D 1H-1H NOESY	1.2 mM protein, 20 mM TCEP, methanol	20	methanol	4.0	ambient	?

NMR Spectrometer Information

Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	600

NMR Refinement

Method	Details	Software
simulated annealing	INITIAL STRUCTURE CALCULATION WAS CARRIED OUT USING DEFAULT SCRIPT ANNEAL_NORDC.PY AND TEMPERATURE SCHEDULE 3500-TO-25 IN STEPS OF 12.5. REFINEMENT WAS PERFORMED FOR 3000 STRUCTURES USING DEFAULT SCRIPT REFINE_NORDC.PY WITH TEMPERATURE SCHEDULES 1500-TO-25 IN STEPS OF 3.25 AND 500-TO-25 IN STEPS OF 2.25. FOR STRONGER J-COUPLING CONSTRAINTS ANNEALING DURING REFINEMENT ITS SCALING FACTOR IS RAMPED FROM 1 TO 3 DURING COOLING: JCOUP = CREATE_JCOUPPOT("JCOUP","JNA_COUP_SPBC_IN_METHANOL9.TBL", A=6.98,B=-1.38,C=1.72,PHASE=-60.0) RAMPEDPARAMS.APPEND( MULTRAMP(1,3, "JCOUP.SETSCALE( VALUE )") )	1

NMR Ensemble Information

Conformer Selection Criteria	structures with favorable non-bond energy
Conformers Calculated Total Number	3000
Conformers Submitted Total Number	10
Representative Model	1 (closest to the average)

Computation: NMR Software

#	Classification	Version	Software Name	Author
1	data analysis	?	NMRPipe	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
2	processing	?	NMRPipe	Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
3	data analysis	?	Sparky	Goddard
4	chemical shift assignment	?	Sparky	Goddard
5	peak picking	?	Sparky	Goddard
6	structure solution	2.32	X-PLOR NIH	Schwieters, Kuszewski, Tjandra and Clore
7	refinement	2.32	X-PLOR NIH	Schwieters, Kuszewski, Tjandra and Clore
8	collection	?	TopSpin	Bruker Biospin
9	data analysis	?	TopSpin	Bruker Biospin
10	geometry optimization	?	ProcheckNMR	Laskowski and MacArthur