5JYV

NMR structure of foldswitch-stablized KaiB in complex with pseudo receiver domain of CikA from Thermosynechococcus elongatus

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	3D HNCACB	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
2	3D HN(COCA)CB	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
3	3D HNCO	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
4	3D HN(CA)CO	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
5	3D HBHA(CO)NH	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
16	3D 15N edited 1H-15N NOESY	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
15	3D 13C edited 1H-13C NOESY	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
14	3D HCCH-COSY	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
13	3D HCCH-TOCSY	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
12	3D IPAP-HNCO(CA)	400 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 440 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
11	3D IPAP-HNCO(CA)	650 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 715 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
10	3D IPAP-HNCO	400 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 440 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
9	3D IPAP-HNCO	650 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 715 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
8	2D IPAP-1H-15N HSQC	400 uM [U-99% 15N] pseudo receiver domain of CikA, 440 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
7	2D IPAP-1H-15N HSQC	400 uM [U-99% 15N] pseudo receiver domain of CikA, 440 uM unlabeled KaiB mutant-N29A	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
6	15N, 13C edited 4D NOESY-HSQC	800 uM [U-100% 15N] pseudo receiver domain of CikA, 880 uM [U-100% 13C] KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
45	13C-edited, 12C-filtered 3D NOESY-HSQC	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
44	3D HNCACB	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
43	3D HN(COCA)CB	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
42	3D HNCO	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
41	3D HN(CA)CO	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
40	3D HBHA(CO)NH	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
39	3D 15N edited 1H-15N NOESY	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
38	3D 13C edited 1H-13C NOESY	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
37	3D HCCH-COSY	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
36	3D HCCH-TOCSY	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
35	3D HCAN	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
34	3D HCA(CO)N	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
33	3D IPAP-HNCO(CA)	400 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 440 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
32	3D IPAP-HNCO(CA)	700 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 770 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
31	3D IPAP-HNCO	400 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 440 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
30	3D IPAP-HNCO	700 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 770 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
29	2D IPAP-1H-15N HSQC	400 uM [U-99% 15N] KaiB mutant-N29A, 440 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
28	2D IPAP-1H-15N HSQC	400 uM [U-99% 15N] KaiB mutant-N29A, 440 uM unlabeled pseudo receiver domain of CikA	90% H2O/10% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
27	15N, 13C edited 4D NOESY-HSQC	800 uM [U-100% 15N] KaiB mutant-N29A, 880 uM [U-100% 13C] pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
26	13C-edited, 12C-filtered 3D NOESY-HSQC	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
47	2D 1H-15N HSQC	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
46	2D 1H-15N HSQC	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	95% H2O/5% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
49	2D 1H-13C HSQC	800 uM [U-99% 13C; U-99% 15N] pseudo receiver domain of CikA, 880 uM unlabeled KaiB mutant-N29A	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600
48	2D 1H-13C HSQC	800 uM [U-99% 13C; U-99% 15N] KaiB mutant-N29A, 880 uM unlabeled pseudo receiver domain of CikA	100% D2O	0.1 mM	7	1 atm	50	Bruker AVANCE III 600

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE III	600

NMR Refinement
Method	Details	Software
simulated annealing		X-PLOR NIH

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
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	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore
2	peak picking	PIPP		Garrett
3	processing	NMRPipe		Delaglio, Grzesiek, Vuister, Zhu, Pfeifer and Bax
4	peak picking	Xipp		Garrett DS, Powers R, Gronenborn AM, Clore GM. J Magn Reson. 2011 Dec;213(2):357-63. doi: 10.1016/j.jmr.2011.09.007. A common sense approach to peak picking in two-, three-, and four-dimensional spectra using automatic computer analysis of contour diagrams http://spin.niddk.nih.gov/dgarrett/Xipp/xipp.html
5	chemical shift assignment	MARS		Mars - robust automatic backbone assignment of proteins Journal of Biomolecular NMR, 2004, Volume 30, Number 1, Page 11 Young-Sang Jung, Markus Zweckstetter
6	structure calculation	X-PLOR NIH		Schwieters, Kuszewski, Tjandra and Clore

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.

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5JYV

NMR structure of foldswitch-stablized KaiB in complex with pseudo receiver domain of CikA from Thermosynechococcus elongatus