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BMRB: 30529

6MV3

NMR structure of the cNTnC-cTnI chimera bound to calcium desensitizer W7

Authors

Cai, F., Hwang, P.M., Sykes, B.D.

Assembly

Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle

Entity

1. Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle, entity 1 (polymer, Thiol state: not present), 125 monomers, 14162.94 Da Detail

MDDIYKAAVE QLTEEQKNEF KAAFDIFVLG AEDGSISTKE LGKVMRMLGQ NPTPEELQEM IDEVDEDGSG TVDFDEFLVM MVRSMKDDSK GKFKRPTLRR VRISADAMMQ ALLGARAKGH HHHHH

2. Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle, entity CA (non-polymer), 40.078 Da
3. Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle, entity WW7 (non-polymer), 340.868 Da

Total weight

14543.887 Da

Max. entity weight

14162.94 Da

Source organism

Homo sapiens

Exptl. method

solution NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 96.0 %, Completeness: 81.4 %, Completeness (bb): 79.7 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	81.4 % (1176 of 1445)	86.8 % (656 of 756)	72.5 % (406 of 560)	88.4 % (114 of 129)
Backbone	79.7 % (593 of 744)	92.6 % (237 of 256)	66.1 % (242 of 366)	93.4 % (114 of 122)
Sidechain	84.7 % (692 of 817)	83.8 % (419 of 500)	88.1 % (273 of 310)	0.0 % (0 of 7)
Aromatic	42.4 % (39 of 92)	43.5 % (20 of 46)	41.3 % (19 of 46)
Methyl	97.5 % (115 of 118)	94.9 % (56 of 59)	100.0 % (59 of 59)

1. entity 1

MDDIYKAAVE QLTEEQKNEF KAAFDIFVLG AEDGSISTKE LGKVMRMLGQ NPTPEELQEM IDEVDEDGSG TVDFDEFLVM MVRSMKDDSK GKFKRPTLRR VRISADAMMQ ALLGARAKGH HHHHH

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 303 K, pH 6.8, Details 0.5 mM [U-15N] cChimera_protein, 100 mM potassium chloride, 10 mM imidazole, 10 mM calcium chloride, 0.25 mM [U-99% 2H] DSS, 2.5 mM WW7, 1 % [U-99% 2H] DMSO, 95% H2O/5% D2O

#	Name	Isotope labeling	Concentration
1	cChimera_protein	[U-15N]	0.5 mM
2	potassium chloride	natural abundance	100 mM
3	imidazole	natural abundance	10 mM
4	calcium chloride	natural abundance	10 mM
5	DSS	[U-99% 2H]	0.25 mM
6	WW7	natural abundance	2.5 mM
7	DMSO	[U-99% 2H]	1 %

Sample #2

Solvent system 100% D2O, Pressure 1 atm, Temperature 303 K, pH 6.8, Details 0.5 mM [U-13C; U-15N] cChimera_protein, 100 mM potassium chloride, 10 mM imidazole, 10 mM calcium chloride, 0.25 mM [U-99% 2H] DSS, 2.5 mM WW7, 1 % [U-99% 2H] DMSO, 100% D2O

#	Name	Isotope labeling	Concentration
8	cChimera_protein	[U-13C; U-15N]	0.5 mM
9	potassium chloride	natural abundance	100 mM
10	imidazole	natural abundance	10 mM
11	calcium chloride	natural abundance	10 mM
12	DSS	[U-99% 2H]	0.25 mM
13	WW7	natural abundance	2.5 mM
14	DMSO	[U-99% 2H]	1 %

LACS Plot; CA

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

Referencing offset: -0.05 ppm, Outliers: 1 Detail

LACS Plot; CB

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

Referencing offset: -0.05 ppm, Outliers: 1 Detail

LACS Plot; HA

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

Referencing offset: 0.01 ppm, Outliers: 3 Detail

Release date

2018-11-04

Citation

Structural Changes Induced by the Binding of the Calcium Desensitizer W7 to Cardiac Troponin
Cai, F., Hwang, P.M., Sykes, B.D.
Biochemistry (2018), 57, 6461-6469, PubMed 30376637 , DOI 10.1021/acs.biochem.8b00882 , Abstract

Related entities 1. Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle, entity 1, : 1 : 4 : 4 : 40 entities Detail

Experiments performed 6 experiments Detail

1 2D 1H-15N HSQC

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 303 K, pH 6.8, Details 0.5 mM [U-15N] cChimera_protein, 100 mM potassium chloride, 10 mM imidazole, 10 mM calcium chloride, 0.25 mM [U-99% 2H] DSS, 2.5 mM WW7, 1 % [U-99% 2H] DMSO, 95% H2O/5% D2O

#	Name	Isotope labeling	Concentration
1	cChimera_protein	[U-15N]	0.5 mM
2	potassium chloride	natural abundance	100 mM
3	imidazole	natural abundance	10 mM
4	calcium chloride	natural abundance	10 mM
5	DSS	[U-99% 2H]	0.25 mM
6	WW7	natural abundance	2.5 mM
7	DMSO	[U-99% 2H]	1 %

2 2D 1H-13C HSQC

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 303 K, pH 6.8, Details 0.5 mM [U-13C; U-15N] cChimera_protein, 100 mM potassium chloride, 10 mM imidazole, 10 mM calcium chloride, 0.25 mM [U-99% 2H] DSS, 2.5 mM WW7, 1 % [U-99% 2H] DMSO, 100% D2O

#	Name	Isotope labeling	Concentration
8	cChimera_protein	[U-13C; U-15N]	0.5 mM
9	potassium chloride	natural abundance	100 mM
10	imidazole	natural abundance	10 mM
11	calcium chloride	natural abundance	10 mM
12	DSS	[U-99% 2H]	0.25 mM
13	WW7	natural abundance	2.5 mM
14	DMSO	[U-99% 2H]	1 %

3 3D 1H-15N NOESY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	cChimera_protein	[U-15N]	0.5 mM
2	potassium chloride	natural abundance	100 mM
3	imidazole	natural abundance	10 mM
4	calcium chloride	natural abundance	10 mM
5	DSS	[U-99% 2H]	0.25 mM
6	WW7	natural abundance	2.5 mM
7	DMSO	[U-99% 2H]	1 %

4 3D 1H-13C NOESY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
8	cChimera_protein	[U-13C; U-15N]	0.5 mM
9	potassium chloride	natural abundance	100 mM
10	imidazole	natural abundance	10 mM
11	calcium chloride	natural abundance	10 mM
12	DSS	[U-99% 2H]	0.25 mM
13	WW7	natural abundance	2.5 mM
14	DMSO	[U-99% 2H]	1 %

5 2D TOCSY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
8	cChimera_protein	[U-13C; U-15N]	0.5 mM
9	potassium chloride	natural abundance	100 mM
10	imidazole	natural abundance	10 mM
11	calcium chloride	natural abundance	10 mM
12	DSS	[U-99% 2H]	0.25 mM
13	WW7	natural abundance	2.5 mM
14	DMSO	[U-99% 2H]	1 %

6 2D 13C,15N-double-filtered NOESY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
8	cChimera_protein	[U-13C; U-15N]	0.5 mM
9	potassium chloride	natural abundance	100 mM
10	imidazole	natural abundance	10 mM
11	calcium chloride	natural abundance	10 mM
12	DSS	[U-99% 2H]	0.25 mM
13	WW7	natural abundance	2.5 mM
14	DMSO	[U-99% 2H]	1 %

NMR combined restraints 3 contents Detail

Properties

Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints	combined_30529_6mv3.nef
Input source #2: Coordindates	6mv3.cif
Diamagnetism of the molecular assembly	False (excluding Oxygen atoms)
Whether the assembly has a disulfide bond	None
Whether the assembly has a other bond	True (see coodinates for details)
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)

Ptnr_site_1	Ptnr_site_2	Redox_state_prediction_1	Redox_state_prediction_2	Distance (Å)
1:65:ASP:OD1	2:1:CA:CA	unknown	unknown	n/a
1:67:ASP:OD1	2:1:CA:CA	unknown	unknown	n/a
1:67:ASP:OD2	2:1:CA:CA	unknown	unknown	n/a
1:69:SER:OG	2:1:CA:CA	unknown	unknown	n/a
1:71:THR:O	2:1:CA:CA	unknown	unknown	n/a
1:76:GLU:OE1	2:1:CA:CA	unknown	unknown	n/a
1:76:GLU:OE2	2:1:CA:CA	unknown	unknown	n/a

Non-standard residues

Chain_ID	Seq_ID	Comp_ID	Chem_comp_name	Experimental evidences
B	1	WW7	N-(6-AMINOHEXYL)-5-CHLORO-1-NAPHTHALENESULFONAMIDE	None
C	1	CA	CALCIUM ION	None

Sequence alignments

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

--------10--------20--------30--------40--------50--------60--------70--------80--------90--140-----
MDDIYKAAVEQLTEEQKNEFKAAFDIFVLGAEDGSISTKELGKVMRMLGQNPTPEELQEMIDEVDEDGSGTVDFDEFLVMMVRSMKDDSKGKFKRPTLRR
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
MDDIYKAAVEQLTEEQKNEFKAAFDIFVLGAEDGSISTKELGKVMRMLGQNPTPEELQEMIDEVDEDGSGTVDFDEFLVMMVRSMKDDSKGKFKRPTLRR
--------10--------20--------30--------40--------50--------60--------70--------80--------90-------100

--150-------160-------170
VRISADAMMQALLGARAKGHHHHHH
|||||||||||||||||||||||||
VRISADAMMQALLGARAKGHHHHHH
-------110-------120-----

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	125	0	0	100.0

Content subtype: combined_30529_6mv3.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Covalent bonds	assembly	OK	7		No information
1	Assigned chemical shifts	assigned_chemical_shifts_1	Warning	1170		96.0 (chain: A, length: 125)
1	Distance restraints	CNS/XPLOR_distance_constraints_2	OK	2066 (1)	noe	92.0 (chain: A, length: 125)

Assigned chemical shifts

Saveframe: assigned_chemical_shifts_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 125, Sequence coverage: 96.0%
- Total number of assignments
  - ¹H chemical shifts: 665
  - ¹³C chemical shifts: 391
  - ¹⁵N chemical shifts: 114
- Completeness of assignments
  - Entity_assemble_ID: A
- 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

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	756	664	87.8
¹³C chemical shifts	560	391	69.8
¹⁵N chemical shifts	136	114	83.8

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	256	242	94.5
¹³C chemical shifts	250	119	47.6
¹⁵N chemical shifts	122	114	93.4

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	500	422	84.4
¹³C chemical shifts	310	272	87.7
¹⁵N chemical shifts	14	0	0.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	68	68	100.0
¹³C chemical shifts	68	68	100.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	46	18	39.1
¹³C chemical shifts	46	17	37.0

Covalent bonds

Saveframe: assembly
- Status: OK

Distance restraints

Saveframe: CNS/XPLOR_distance_constraints_2
- Status: OK
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 125, Sequence coverage: 92.0%
- Total number of restraints: 2066
- Weight of restraints: 1.0 (2066)
- Potential type of restraints: square-well-parabolic (2066)
- Range of distance target values: 1.57, 4.36 (Å)
- Histogram of distance target values
- Histogram of discrepancy in distance target values
- Distance restraints per residue
  - Chain_ID: A
  - Chain_ID: B
    None
  - Chain_ID: C
    None
- Distance restraints on contact map

Keywords EF hand, METAL BINDING PROTEIN, W7, calcium binding protein, calcium desensitizer, cardiac troponin

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Found 1 Document (0.528 seconds)

BMRB: 30529

Sample #1

Sample #2

Chemical shift reference

Chemical shift reference

Chemical shift reference

Related entities 1. Troponin C, slow skeletal and cardiac muscles,Troponin I, cardiac muscle, entity 1, : 1 : 4 : 4 : 40 entities Detail

Experiments performed 6 experiments Detail

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

Instrument

Sample

NMR combined restraints 3 contents Detail