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BMRB: 18027

2LL6

Solution NMR structure of CaM bound to iNOS CaM binding domain peptide

Authors

Piazza, M., Futrega, K., Spratt, D.E., Dieckmann, T., Guillemette, J.GUY.

Assembly

CaM bound to NOS peptides

Entity

1. CaM (polymer, Thiol state: all free), 148 monomers, 16706.21 Da Detail

ADQLTEEQIA EFKEAFSLFD KDGDGTITTK ELGTVMRSLG QNPTEAELQD MINEVDADGN GTIDFPEFLT MMARKMKDTD SEEEIREAFR VFDKDGNGYI SAAELRHVMT NLGEKLTDEE VDEMIREADI DGDGQVNYEE FVQMMTAK

2. iNOS (polymer), 17 monomers, 1963.603 Da Detail

LKVLVKAVLF ACMLMRK

Total weight

18669.814 Da

Max. entity weight

16706.21 Da

Source organism

Exptl. method

Refine. method

DGSA-distance geometry simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 98.2 %, Completeness: 76.4 %, Completeness (bb): 80.6 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	76.4 % (1445 of 1891)	85.2 % (837 of 982)	60.6 % (445 of 734)	93.1 % (163 of 175)
Backbone	80.6 % (795 of 986)	97.6 % (331 of 339)	62.4 % (302 of 484)	99.4 % (162 of 163)
Sidechain	73.7 % (780 of 1059)	78.7 % (506 of 643)	67.6 % (273 of 404)	8.3 % (1 of 12)
Aromatic	11.8 % (13 of 110)	23.6 % (13 of 55)	0.0 % (0 of 55)
Methyl	83.3 % (145 of 174)	92.0 % (80 of 87)	74.7 % (65 of 87)

1. CaM

ADQLTEEQIA EFKEAFSLFD KDGDGTITTK ELGTVMRSLG QNPTEAELQD MINEVDADGN GTIDFPEFLT MMARKMKDTD SEEEIREAFR VFDKDGNGYI SAAELRHVMT NLGEKLTDEE VDEMIREADI DGDGQVNYEE FVQMMTAK

2. iNOS

LKVLVKAVLF ACMLMRK

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

Protein Blocks Logo

Calculated from 20 models in PDB: 2LL6, Strand ID: A, B Detail

Release date

2012-05-14

Citation

Structure and dynamics of calmodulin (CaM) bound to nitric oxide synthase peptides: effects of a phosphomimetic CaM mutation
Piazza, M., Futrega, K., Spratt, D.E., Dieckmann, T., Guillemette, J.GUY.
Biochemistry (2012), 51, 3651-3661, PubMed 22486744 , DOI 10.1021/bi300327z , Abstract

Related entities 1. CaM, : 11 : 88 entities Detail

Related entities 2. iNOS, : 1 : 4 : 1 : 6 entities Detail

Interaction partners 2. iNOS, : 5 interactors Detail

More details for 5 interactors identified by 2 citations

Experiments performed 14 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

2 2D 1H-15N HSQC

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

3 3D CBCA(CO)NH

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

4 3D CBCA(CO)NH

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

5 3D HNCA

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

6 3D HNCO

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

7 3D HBHA(CO)NH

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

8 3D HNCA

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

9 3D H(CCO)NH

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

10 3D HCCH-TOCSY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

11 3D 1H-15N NOESY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

12 3D 1H-15N NOESY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

13 3D HCCH-COSY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-99% 13C; U-99% 15N]		1.0 mM
2	entity_2	natural abundance		1.0 mM
3	potassium chloride	natural abundance		100 mM
4	calcium chloride	natural abundance		10 mM
5	sodium azide	natural abundance		0.2 mM
6	H2O	natural abundance		90 %
7	D2O	natural abundance		10 %

14 3D HCCH-COSY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.0

#	Name	Isotope labeling	Type	Concentration
8	entity_1	natural abundance		1.0 mM
9	entity_2	[U-99% 13C; U-99% 15N]		1.0 mM
10	potassium chloride	natural abundance		100 mM
11	calcium chloride	natural abundance		10 mM
12	sodium azide	natural abundance		0.2 mM
13	H2O	natural abundance		90 %
14	D2O	natural abundance		10 %

NMR combined restraints 4 contents Detail

Properties

Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints	combined_18027_2ll6.nef
Input source #2: Coordindates	2ll6.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--------90-------100
ADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYI
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYI

-------110-------120-------130-------140--------
SAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK
||||||||||||||||||||||||||||||||||||||||||||||||
SAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK

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

-150-----160-----
LKVLVKAVLFACMLMRK
|||||||||||||||||
LKVLVKAVLFACMLMRK
--------10-------

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	148	0	0	100.0
B	B	17	0	0	100.0

Content subtype: combined_18027_2ll6.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chem_shift_list_1	Warning	1438		98.0 (chain: A, length: 148), 100.0 (chain: B, length: 17)
1	Distance restraints	CNS/XPLOR_distance_constraints_2	Warning	3434 (1)	noe	97.3 (chain: A, length: 148), 100.0 (chain: B, length: 17)
1	Dihedral angle restraints	CNS/XPLOR_dihedral_3	OK	281 (281)	.	98.0 (chain: A, length: 148), 52.9 (chain: B, length: 17)

Assigned chemical shifts

Saveframe: assigned_chem_shift_list_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 148, Sequence coverage: 98.0%
  - Entity_assembly_ID: B, Chain length: 17, Sequence coverage: 100.0%
- Total number of assignments
  - ¹H chemical shifts: 845
  - ¹³C chemical shifts: 431
  - ¹⁵N chemical shifts: 162
- Completeness of assignments
  - Entity_assemble_ID: A
  - Entity_assemble_ID: B
- Redox state of CYS
- 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

Distance restraints

Saveframe: CNS/XPLOR_distance_constraints_2
- Status: Warning
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 148, Sequence coverage: 97.3%
  - Entity_assembly_ID: B, Chain length: 17, Sequence coverage: 100.0%
- Total number of restraints: 3429
- Weight of restraints: 1.0 (3429)
- Potential type of restraints: square-well-parabolic (3429)
- Range of distance target values: 2.35, 3.65 (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

Dihedral angle restraints

Saveframe: CNS/XPLOR_dihedral_3
- Status: OK
- Experiment type: .
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 148, Sequence coverage: 98.0%
  - Entity_assembly_ID: B, Chain length: 17, Sequence coverage: 52.9%
- Total number of restraints: 281
- Total number of restraints per polymer type: 281
- Weight of restraints: 1.0 (281)
- Potential type of restraints: square-well-parabolic (281)
- Plot of Phi-Psi angle restraints
- Dihedral angle restraints per residue
  - Chain_ID: A
  - Chain_ID: B

Keywords Protein