2MKI

Solution structure of tandem RRM domains of cytoplasmic polyadenylation element binding protein 4 (CPEB4) in complex with RNA

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D 1H-15N HSQC	0.4-0.6 mM [U-100% 15N] CPEB4RRM12-21, 0.4-0.6 mM 5'-CUUUA-3'-22, 100 mM sodium chloride-23, 1 mM DTT-24, 50 mM potassium phosphate-25	90% H2O/10% D2O	0.15	7.0	ambient	303
2	2D 1H-13C HSQC	0.4-0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-6, 0.4-06 mM RNA (5'-CUUUA)-3')-7, 100 mM sodium chloride-8, 1 mM DTT-9, 50 mM sodium phosphate-10	100% D2O	0.15	7.0	ambient	303
3	2D 1H-13C HSQC aliphatic	0.4-0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-6, 0.4-06 mM RNA (5'-CUUUA)-3')-7, 100 mM sodium chloride-8, 1 mM DTT-9, 50 mM sodium phosphate-10	100% D2O	0.15	7.0	ambient	303
4	2D 1H-13C HSQC aromatic	0.4-0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-6, 0.4-06 mM RNA (5'-CUUUA)-3')-7, 100 mM sodium chloride-8, 1 mM DTT-9, 50 mM sodium phosphate-10	100% D2O	0.15	7.0	ambient	303
5	2D 1H-1H TOCSY	0.4-0.6 mM [U-100% 15N] CPEB4RRM12-21, 0.4-0.6 mM 5'-CUUUA-3'-22, 100 mM sodium chloride-23, 1 mM DTT-24, 50 mM potassium phosphate-25	90% H2O/10% D2O	0.15	7.0	ambient	303
6	2D 1H-1H NOESY	0.4-0.6 mM [U-100% 15N] CPEB4RRM12-16, 0.4-0.6 mM 5'-CUUUA-3'-17, 100 mM sodium chloride-18, 1 mM DTT-19, 50 mM potassium phosphate-20	100% D2O	0.15	7.0	ambient	303
7	3D trHNCA	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
8	3D trHN(CO)CA	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
9	3D trCBCA(CO)NH	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
10	3D trHNCACB	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
11	3D trHNCO	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
12	3D trHCACO	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
13	3D HCCH-TOCSY	50 mM sodium phosphate-1, 100 mM sodium chloride-2, 1 mM DTT-3, 0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-4, 0.6 mM 5'-CUUUA-3'-5	90% H2O/10% D2O	0.15	7.0	ambient	303
14	3D 1H-15N NOESY	0.4-0.6 mM [U-100% 15N] CPEB4RRM12-21, 0.4-0.6 mM 5'-CUUUA-3'-22, 100 mM sodium chloride-23, 1 mM DTT-24, 50 mM potassium phosphate-25	90% H2O/10% D2O	0.15	7.0	ambient	303
15	3D 1H-13C NOESY aliphatic	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
16	3D 1H-13C NOESY aromatic	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
17	3D HCCH-TOCSY	0.4-0.6 mM [U-100% 13C; U-100% 15N; U-80% 2H] CPEB4RRM12-11, 0.4-0.6 mM 5'-CUUUA-3'-12, 100 mM sodium chloride-13, 1 mM DTT-14, 50 mM sodium phosphate-15	90% H2O/10% D2O	0.15	7.0	ambient	303
18	2D 1H-15N TROSY	0.4-0.6 mM [U-100% 15N] CPEB4RRM12-21, 0.4-0.6 mM 5'-CUUUA-3'-22, 100 mM sodium chloride-23, 1 mM DTT-24, 50 mM potassium phosphate-25	90% H2O/10% D2O	0.15	7.0	ambient	303
19	2D 13C F1-filtered F2-filtered NOESY	50 mM sodium phosphate-1, 100 mM sodium chloride-2, 1 mM DTT-3, 0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-4, 0.6 mM 5'-CUUUA-3'-5	90% H2O/10% D2O	0.15	7.0	ambient	303
20	2D 1H-1H F1-13C-filtered F2-13C-edited NOESY	50 mM sodium phosphate-1, 100 mM sodium chloride-2, 1 mM DTT-3, 0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-4, 0.6 mM 5'-CUUUA-3'-5	90% H2O/10% D2O	0.15	7.0	ambient	303
21	3D 13C F1-edited, F3-filtered NOESY HSQC	50 mM sodium phosphate-1, 100 mM sodium chloride-2, 1 mM DTT-3, 0.6 mM [U-100% 13C; U-100% 15N] CPEB4RRM12-4, 0.6 mM 5'-CUUUA-3'-5	90% H2O/10% D2O	0.15	7.0	ambient	303

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Bruker	AVANCE	900
2	Bruker	AVANCE	700
3	Bruker	AVANCE	600
4	Bruker	AVANCE	500

NMR Refinement
Method	Details	Software
simulated annealing		Amber

NMR Ensemble Information
Conformer Selection Criteria	structures with the lowest energy
Conformers Calculated Total Number	30
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average)

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	refinement	Amber		Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... and Kollman

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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2MKI

Solution structure of tandem RRM domains of cytoplasmic polyadenylation element binding protein 4 (CPEB4) in complex with RNA