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BMRB: 18314

2LQH

NMR structure of FOXO3a transactivation domains (CR2C-CR3) in complex with CBP KIX domain (2b3l conformation)

Authors

Wang, F., Marshall, C.B., Yamamoto, K., Li, G., Gasmi-Seabrook, G.M. C., Okada, H., Mak, T.W., Ikura, M.

Assembly

FOXO3a

Entity

1. FOXO3a, entity 1 (polymer, Thiol state: all free), 87 monomers, 10331.68 Da Detail

GVRKGWHEHV TQDLRSHLVH KLVQAIFPTP DPAALKDRRM ENLVAYAKKV EGDMYESANS RDEYYHLLAE KIYKIQKELE EKRRSRL

2. FOXO3a, entity 2 (polymer, Thiol state: all free), 52 monomers, 5205.493 Da Detail

GSMSHYGNQT LQDLLTSDSL SHSDGGGSGG GSGGGSLECD MESIIRSELM DA

Total weight

15537.173 Da

Max. entity weight

10331.68 Da

Source organism

Mus musculus , Homo sapiens

Exptl. method

solution NMR

Refine. method

simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 92.8 %, Completeness: 86.8 %, Completeness (bb): 88.2 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	86.8 % (1365 of 1572)	88.4 % (727 of 822)	84.5 % (511 of 605)	87.6 % (127 of 145)
Backbone	88.2 % (730 of 828)	86.5 % (250 of 289)	89.8 % (362 of 403)	86.8 % (118 of 136)
Sidechain	86.8 % (754 of 869)	89.5 % (477 of 533)	82.0 % (268 of 327)	100.0 % (9 of 9)
Aromatic	26.5 % (26 of 98)	51.0 % (25 of 49)	0.0 % (0 of 48)	100.0 % (1 of 1)
Methyl	100.0 % (128 of 128)	100.0 % (64 of 64)	100.0 % (64 of 64)

1. entity 1

GVRKGWHEHV TQDLRSHLVH KLVQAIFPTP DPAALKDRRM ENLVAYAKKV EGDMYESANS RDEYYHLLAE KIYKIQKELE EKRRSRL

2. entity 2

GSMSHYGNQT LQDLLTSDSL SHSDGGGSGG GSGGGSLECD MESIIRSELM DA

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

Sample #2

Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
8	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
9	FOXO3a	natural abundance	0.6 mM
10	MES	natural abundance	20 mM
11	sodium chloride	natural abundance	50 mM
12	DTT	natural abundance	1 mM
13	D2O	natural abundance	100 %

Sample #3

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

Sample #4

Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
21	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
22	KIX	natural abundance	0.6 mM
23	MES	natural abundance	20 mM
24	sodium chloride	natural abundance	50 mM
25	DTT	natural abundance	1 mM
26	D2O	natural abundance	100 %

LACS Plot; CA

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	na	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	na	indirect	0.1013291

Referencing offset: 2.66 ppm, Outliers: 1 Detail

LACS Plot; CB

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	na	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	na	indirect	0.1013291

Referencing offset: 2.66 ppm, Outliers: 1 Detail

LACS Plot; HA

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	na	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	na	indirect	0.1013291

Referencing offset: -0.04 ppm, Outliers: 1 Detail

LACS Plot; CO

Atom type	Mol. common name	Atom group	Ref. method	Ref. type	Shift ratio
¹³C	DSS	methyl protons	na	indirect	0.2514495
¹H	DSS	methyl protons	internal	direct	1.0
¹⁵N	DSS	methyl protons	na	indirect	0.1013291

Referencing offset: 3.02 ppm, Outliers: 3 Detail

Protein Blocks Logo

Calculated from 20 models in PDB: 2LQH, Strand ID: A, B Detail

Release date

2012-05-07

Citation

Structures of KIX domain of CBP in complex with two FOXO3a transactivation domains reveal promiscuity and plasticity in coactivator recruitment
Wang, F., Marshall, C.B., Yamamoto, K., Li, G., Gasmi-Seabrook, G.M.C., Okada, H., Mak, T.W., Ikura, M.
Proc. Natl. Acad. Sci. U. S. A. (2012), 109, 6078-6083, PubMed 22474372 , DOI 10.1073/pnas.1119073109 , Abstract

Related entities 1. FOXO3a, entity 1, : 6 : 23 entities Detail

Related entities 2. FOXO3a, entity 2, : 1 : 1 entities Detail

Interaction partners 1. FOXO3a, entity 1, : 1 interactors Detail

More details for 1 interactors identified by 1 citations

Experiments performed 16 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

2 3D HNCO

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

3 3D CBCA(CO)NH

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

4 3D HNCACB

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

5 3D 1H-15N NOESY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
1	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
2	FOXO3a	natural abundance	0.6 mM
3	MES	natural abundance	20 mM
4	sodium chloride	natural abundance	50 mM
5	DTT	natural abundance	1 mM
6	H2O	natural abundance	90 %
7	D2O	natural abundance	10 %

6 2D 1H-13C HSQC

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
8	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
9	FOXO3a	natural abundance	0.6 mM
10	MES	natural abundance	20 mM
11	sodium chloride	natural abundance	50 mM
12	DTT	natural abundance	1 mM
13	D2O	natural abundance	100 %

7 3D HCCH-TOCSY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
8	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
9	FOXO3a	natural abundance	0.6 mM
10	MES	natural abundance	20 mM
11	sodium chloride	natural abundance	50 mM
12	DTT	natural abundance	1 mM
13	D2O	natural abundance	100 %

8 3D 1H-13C NOESY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
8	KIX	[U-100% 13C; U-100% 15N]	0.6 mM
9	FOXO3a	natural abundance	0.6 mM
10	MES	natural abundance	20 mM
11	sodium chloride	natural abundance	50 mM
12	DTT	natural abundance	1 mM
13	D2O	natural abundance	100 %

9 2D 1H-15N HSQC

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

10 3D HNCO

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

11 3D HNCACB

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

12 3D CBCA(CO)NH

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

13 3D 1H-15N NOESY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
14	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
15	KIX	natural abundance	0.6 mM
16	MES	natural abundance	20 mM
17	sodium chloride	natural abundance	50 mM
18	DTT	natural abundance	1 mM
19	H2O	natural abundance	90 %
20	D2O	natural abundance	10 %

14 2D 1H-13C HSQC

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
21	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
22	KIX	natural abundance	0.6 mM
23	MES	natural abundance	20 mM
24	sodium chloride	natural abundance	50 mM
25	DTT	natural abundance	1 mM
26	D2O	natural abundance	100 %

15 3D HCCH-TOCSY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
21	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
22	KIX	natural abundance	0.6 mM
23	MES	natural abundance	20 mM
24	sodium chloride	natural abundance	50 mM
25	DTT	natural abundance	1 mM
26	D2O	natural abundance	100 %

16 3D 1H-13C NOESY

Instrument

Bruker Avance - 800 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 273 K, pH 6

#	Name	Isotope labeling	Concentration
21	FOXO3a3	[U-100% 13C; U-100% 15N]	0.6 mM
22	KIX	natural abundance	0.6 mM
23	MES	natural abundance	20 mM
24	sodium chloride	natural abundance	50 mM
25	DTT	natural abundance	1 mM
26	D2O	natural abundance	100 %

NMR combined restraints 5 contents Detail

Properties

Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints	combined_18314_2lqh.nef
Input source #2: Coordindates	2lqh.cif
Diamagnetism of the molecular assembly	True (excluding Oxygen atoms)
Whether the assembly has a disulfide bond	None
Whether the assembly has a other bond	None
Whether the assembly contains a cyclic polymer	None
Overall data processing status	Warning

Disulfide bonds

None

Other bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)

None

Non-standard residues

None

Sequence alignments

Entity_assembly_ID (NMR): A vs Auth_asym_ID (model): A

--------10--------20--------30--------40--------50--------60--------70--------80-------
GVRKGWHEHVTQDLRSHLVHKLVQAIFPTPDPAALKDRRMENLVAYAKKVEGDMYESANSRDEYYHLLAEKIYKIQKELEEKRRSRL
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
GVRKGWHEHVTQDLRSHLVHKLVQAIFPTPDPAALKDRRMENLVAYAKKVEGDMYESANSRDEYYHLLAEKIYKIQKELEEKRRSRL

Entity_assembly_ID (NMR): B vs Auth_asym_ID (model): B

-------110-------120-------130-------140-------150--
GSMSHYGNQTLQDLLTSDSLSHSDGGGSGGGSGGGSLECDMESIIRSELMDA
||||||||||||||||||||||||||||||||||||||||||||||||||||
GSMSHYGNQTLQDLLTSDSLSHSDGGGSGGGSGGGSLECDMESIIRSELMDA
--------10--------20--------30--------40--------50--

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	87	0	0	100.0
B	B	52	0	0	100.0

Content subtype: combined_18314_2lqh.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chem_shift_list_1	Warning	1362		100.0 (chain: A, length: 87), 76.9 (chain: B, length: 52)
1	Distance restraints	CNS/XPLOR_distance_constraints_2	Warning	4047 (1)	noe	100.0 (chain: A, length: 87), 65.4 (chain: B, length: 52)
2	Distance restraints	CNS/XPLOR_distance_constraints_4	OK	118 (1)	hbond	66.7 (chain: A, length: 87), 40.4 (chain: B, length: 52)
1	Dihedral angle restraints	CNS/XPLOR_dihedral_3	OK	200 (200)	.	94.3 (chain: A, length: 87), 61.5 (chain: B, length: 52)

Assigned chemical shifts

Saveframe: assigned_chem_shift_list_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 87, Sequence coverage: 100.0%
  - Entity_assembly_ID: B, Chain length: 52, Sequence coverage: 76.9%
- Total number of assignments
  - ¹H chemical shifts: 726
  - ¹³C chemical shifts: 512
  - ¹⁵N chemical shifts: 124
- Completeness of assignments
  - Entity_assemble_ID: A
  - Entity_assemble_ID: B
- Redox state of CYS
- Peptide bond of PRO
- Rotameric state of ILE , LEU, and VAL
- Histogram of normalized assigned chemical shifts
- Random coil index and derived parameters (S² and NMR RMSD)
  - Chain_ID: A
  - Chain_ID: B

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	564	528	93.6
¹³C chemical shifts	413	365	88.4
¹⁵N chemical shifts	98	87	88.8

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	174	170	97.7
¹³C chemical shifts	174	169	97.1
¹⁵N chemical shifts	84	81	96.4

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	390	358	91.8
¹³C chemical shifts	239	196	82.0
¹⁵N chemical shifts	14	6	42.9

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	47	47	100.0
¹³C chemical shifts	47	47	100.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	41	23	56.1
¹³C chemical shifts	40	0	0.0
¹⁵N chemical shifts	1	1	100.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	258	198	76.7
¹³C chemical shifts	192	147	76.6
¹⁵N chemical shifts	56	37	66.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	115	76	66.1
¹³C chemical shifts	104	73	70.2
¹⁵N chemical shifts	52	34	65.4

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	143	122	85.3
¹³C chemical shifts	88	74	84.1
¹⁵N chemical shifts	4	3	75.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	22	20	90.9
¹³C chemical shifts	22	20	90.9

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	8	0	0.0
¹³C chemical shifts	8	0	0.0

Distance restraints

Saveframe: CNS/XPLOR_distance_constraints_2
- Status: Warning
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 87, Sequence coverage: 100.0%
  - Entity_assembly_ID: B, Chain length: 52, Sequence coverage: 65.4%
- Total number of restraints: 4035
- Weight of restraints: 1.0 (4035)
- Potential type of restraints: square-well-parabolic (4035)
- Range of distance target values: 2.25, 4.05 (Å)
- Histogram of distance target values
- Histogram of discrepancy in distance target values
- Distance restraints per residue
  - Chain_ID: A
  - Chain_ID: B
- Distance restraints on contact map
- Saveframe: CNS/XPLOR_distance_constraints_4
  - Status: OK
  - Experiment type: hbond
  - Sequence coverage of experimental data
    - Entity_assembly_ID: A, Chain length: 87, Sequence coverage: 66.7%
    - Entity_assembly_ID: B, Chain length: 52, Sequence coverage: 40.4%
  - Total number of restraints: 118
  - Weight of restraints: 1.0 (118)
  - Potential type of restraints: square-well-parabolic (118)
  - Range of distance target values: 1.95, 2.9 (Å)
  - Histogram of distance target values
  - Distance restraints per residue
    - Chain_ID: A
    - Chain_ID: B
  - Distance restraints on contact map

Dihedral angle restraints

Saveframe: CNS/XPLOR_dihedral_3
- Status: OK
- Experiment type: .
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 87, Sequence coverage: 94.3%
  - Entity_assembly_ID: B, Chain length: 52, Sequence coverage: 61.5%
- Total number of restraints: 200
- Total number of restraints per polymer type: 200
- Weight of restraints: 1.0 (200)
- Potential type of restraints: square-well-parabolic (200)
- Plot of Phi-Psi angle restraints
- Dihedral angle restraints per residue
  - Chain_ID: A
  - Chain_ID: B

Keywords intrinsic disorder, NMR solution structure, promiscuous binding, transcription

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Found 1 Document (0.835 seconds)

BMRB: 18314

Sample #1

Sample #2

Sample #3

Sample #4

Chemical shift reference

Chemical shift reference

Chemical shift reference

Chemical shift reference

Related entities 1. FOXO3a, entity 1, : 6 : 23 entities Detail

Related entities 2. FOXO3a, entity 2, : 1 : 1 entities Detail

Interaction partners 1. FOXO3a, entity 1, : 1 interactors Detail

Experiments performed 16 experiments Detail

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NMR combined restraints 5 contents Detail