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BMRB: 16944

2KYB

Chemical shift assignment of CpR82G from Clostridium perfringens. North East Structural Genomics Consortium Target CpR82g.

Authors

Mobley, C.K., Lee, H., Lee, D., Ciccosanti, C., Janjua, H., Acton, T.B., Xiao, R., Everrett, J.K., Montelione, G.T., Prestegard, J.H.

Assembly

CpR82G

Entity

1. CpR82G (polymer, Thiol state: not present), 68 monomers, 7396.186 Da Detail

MKTGIVNVSS SLNVRSSAST SSKVIGSLSG NTKVTIVGEE GAFYKIEYKG SHGYVAKEYI LEHHHHHH

Formula weight

7396.186 Da

Source organism

Clostridium perfringens

Exptl. method

solution NMR

Refine. method

simulated annealing

Data set

assigned_chemical_shifts, RDCs

Chem. Shift Complete

Sequence coverage: 91.2 %, Completeness: 78.6 %, Completeness (bb): 90.0 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	78.6 % (597 of 760)	74.2 % (288 of 388)	82.4 % (248 of 301)	85.9 % (61 of 71)
Backbone	90.0 % (367 of 408)	90.2 % (129 of 143)	89.8 % (177 of 197)	89.7 % (61 of 68)
Sidechain	68.8 % (284 of 413)	64.9 % (159 of 245)	75.8 % (125 of 165)	0.0 % (0 of 3)
Aromatic	28.6 % (20 of 70)	28.6 % (10 of 35)	28.6 % (10 of 35)
Methyl	83.8 % (62 of 74)	81.1 % (30 of 37)	86.5 % (32 of 37)

1. CpR82G

MKTGIVNVSS SLNVRSSAST SSKVIGSLSG NTKVTIVGEE GAFYKIEYKG SHGYVAKEYI LEHHHHHH

Show sample condition

Sample #1

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
9	CpR82G	U-5% 13C; U-100% 15N	0.98 mM
10	sodium azide	natural abundance	0.02 %
11	DTT	natural abundance	100 mM
12	Calcium Chloride	natural abundance	5 mM
13	sodium chloride	natural abundance	100 mM
14	MES	natural abundance	20 mM
15	H2O	natural abundance	95 %
16	D2O	natural abundance	5 %

Sample #3

Solvent system 82.88% H2O/17.12% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
17	CpR82G	[U-100% 13C; U-100% 15N]	0.67 mM
18	sodium azide	natural abundance	0.016 %
19	DTT	natural abundance	81.73 mM
20	Calcium Chloride	natural abundance	4.09 mM
21	sodium chloride	natural abundance	81.73 mM
22	MES	natural abundance	16.35 mM
23	C12E5 PEG	natural abundance	4 %
24	Hexanol	natural abundance	1.25 %
25	H2O	natural abundance	82.88 %
26	D2O	natural abundance	17.12 %

Sample #4

Solvent system 91.3% H2O/8.7% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
27	CpR82G	natural abundance	0.92 mM
28	sodium azide	natural abundance	0.018 %
29	DTT	natural abundance	91.67 mM
30	Calcium Chloride	natural abundance	4.58 mM
31	sodium chloride	natural abundance	91.67 mM
32	MES	natural abundance	18.32 mM
33	Positively Charged Compressed Polyacrylamide Gel	natural abundance	7 %
34	H2O	natural abundance	91.3 %
35	D2O	natural abundance	8.7 %

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: -0.1 ppm, Outliers: 2 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: -0.1 ppm, Outliers: 2 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: -0.05 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: 0.76 ppm, Outliers: 3 Detail

Protein Blocks Logo

Calculated from 10 models in PDB: 2KYB, Strand ID: A Detail

RDC

95 RDC values in 2 lists
Field strength (¹H) 600 MHz, Pressure 1 atm, Temperature 298 K, pH 6.5 Detail

Release date

2010-06-21

Citation

Solution Structure of CpR82G
Mobley, C.K., Lee, H., Montelione, G.T., Prestegard, J.H.
Not known

Related entities 1. CpR82G, : 1 : 1 : 9 entities Detail

Experiments performed 19 experiments Detail

1 3D C(CO)NH

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

2 3D 1H-15N NOESY

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

3 3D 1H-13C NOESY

Instrument

Varian INOVA - 600 MHz Withe cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

4 3D CBCA(CO)NH

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

5 2D 1H-13C HSQC

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

6 2D 1H-15N HSQC

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

7 3D HBHA(CO)NH

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

8 3D HCCH-TOCSY

Instrument

Varian INOVA - 600 MHz Withe cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

9 3D H(CCO)NH

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

10 3D HNCACB

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

11 2D 1H-13C HSQC CT 28ms delay

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
9	CpR82G	U-5% 13C; U-100% 15N	0.98 mM
10	sodium azide	natural abundance	0.02 %
11	DTT	natural abundance	100 mM
12	Calcium Chloride	natural abundance	5 mM
13	sodium chloride	natural abundance	100 mM
14	MES	natural abundance	20 mM
15	H2O	natural abundance	95 %
16	D2O	natural abundance	5 %

12 2D 1H-13C HSQC CT 42ms delay

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
9	CpR82G	U-5% 13C; U-100% 15N	0.98 mM
10	sodium azide	natural abundance	0.02 %
11	DTT	natural abundance	100 mM
12	Calcium Chloride	natural abundance	5 mM
13	sodium chloride	natural abundance	100 mM
14	MES	natural abundance	20 mM
15	H2O	natural abundance	95 %
16	D2O	natural abundance	5 %

13 2D 1H-13C HSQC CT 56ms delay

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
9	CpR82G	U-5% 13C; U-100% 15N	0.98 mM
10	sodium azide	natural abundance	0.02 %
11	DTT	natural abundance	100 mM
12	Calcium Chloride	natural abundance	5 mM
13	sodium chloride	natural abundance	100 mM
14	MES	natural abundance	20 mM
15	H2O	natural abundance	95 %
16	D2O	natural abundance	5 %

14 NH-J-modulation

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 82.88% H2O/17.12% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
17	CpR82G	[U-100% 13C; U-100% 15N]	0.67 mM
18	sodium azide	natural abundance	0.016 %
19	DTT	natural abundance	81.73 mM
20	Calcium Chloride	natural abundance	4.09 mM
21	sodium chloride	natural abundance	81.73 mM
22	MES	natural abundance	16.35 mM
23	C12E5 PEG	natural abundance	4 %
24	Hexanol	natural abundance	1.25 %
25	H2O	natural abundance	82.88 %
26	D2O	natural abundance	17.12 %

15 NH-J-modulation

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 91.3% H2O/8.7% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
27	CpR82G	natural abundance	0.92 mM
28	sodium azide	natural abundance	0.018 %
29	DTT	natural abundance	91.67 mM
30	Calcium Chloride	natural abundance	4.58 mM
31	sodium chloride	natural abundance	91.67 mM
32	MES	natural abundance	18.32 mM
33	Positively Charged Compressed Polyacrylamide Gel	natural abundance	7 %
34	H2O	natural abundance	91.3 %
35	D2O	natural abundance	8.7 %

16 3D HNCO

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

17 hbcbcgcdhdA

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

18 hbcbcgcdceheA

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

19 2D 1H-13C HSQC aromatic

Instrument

Varian INOVA - 600 MHz With cold probe

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	CpR82G	[U-100% 13C; U-100% 15N]	1.01 mM
2	sodium azide	natural abundance	0.02 %
3	DTT	natural abundance	100 mM
4	Calcium Chloride	natural abundance	5 mM
5	sodium chloride	natural abundance	100 mM
6	MES	natural abundance	20 mM
7	H2O	natural abundance	95 %
8	D2O	natural abundance	5 %

NMR combined restraints 4 contents Detail

Properties

Input source #1: NMR data (NEF) - Assigned chemical shifts, Distance restraints, Dihedral angle restraints	combined_16944_2kyb.nef
Input source #2: Coordindates	2kyb.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

Chain_ID	Seq_ID	Comp_ID	Chem_comp_name	Experimental evidences
A	61	.	Not matched with CCD	None

Sequence alignments

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

--------10--------20--------30--------40--------50--------60---
MKTGIVNVSSSLNVRSSASTSSKVIGSLSGNTKVTIVGEEGAFYKIEYKGSHGYVAKEYI.EL
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||   
MKTGIVNVSSSLNVRSSASTSSKVIGSLSGNTKVTIVGEEGAFYKIEYKGSHGYVAKEYI   
--------10--------20--------30--------40--------50--------60

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	60	3	0	100.0

Content subtype: combined_16944_2kyb.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chem_shift_list_1	Warning	585		96.8 (chain: A, length: 63)
1	Distance restraints	CNS/XPLOR_distance_constraints_2	Warning	728 (1)	noe	93.7 (chain: A, length: 63)
1	Dihedral angle restraints	CNS/XPLOR_dihedral_3	OK	63 (63)	.	77.8 (chain: A, length: 63)

Assigned chemical shifts

Saveframe: assigned_chem_shift_list_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 63, Sequence coverage: 96.8%
- Total number of assignments
  - ¹H chemical shifts: 281
  - ¹³C chemical shifts: 244
  - ¹⁵N chemical shifts: 60
- Completeness of assignments
  - Entity_assemble_ID: A
- Tautomeric state of HIS
- 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	352	281	79.8
¹³C chemical shifts	271	244	90.0
¹⁵N chemical shifts	66	60	90.9

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	131	127	96.9
¹³C chemical shifts	124	121	97.6
¹⁵N chemical shifts	62	60	96.8

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	221	154	69.7
¹³C chemical shifts	147	123	83.7
¹⁵N chemical shifts	4	0	0.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	38	30	78.9
¹³C chemical shifts	38	31	81.6

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	23	10	43.5
¹³C chemical shifts	23	10	43.5

Distance restraints

Saveframe: CNS/XPLOR_distance_constraints_2
- Status: Warning
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 63, Sequence coverage: 93.7%
- Total number of restraints: 718
- Weight of restraints: 1.0 (718)
- Potential type of restraints: square-well-parabolic (718)
- Range of distance target values: 2.06, 3.9 (Å)
- Histogram of distance target values
- Histogram of discrepancy in distance target values
- Distance restraints per residue
  - Chain_ID: A
- 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: 63, Sequence coverage: 77.8%
- Total number of restraints: 63
- Total number of restraints per polymer type: 63
- Weight of restraints: 1.0 (63)
- Potential type of restraints: square-well-parabolic (63)
- Plot of Phi-Psi angle restraints
- Dihedral angle restraints per residue
  - Chain_ID: A

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Found 1 Document (0.62 seconds)

BMRB: 16944

Sample #1

Sample #2

Sample #3

Sample #4

Chemical shift reference

Chemical shift reference

Chemical shift reference

Chemical shift reference

Related entities 1. CpR82G, : 1 : 1 : 9 entities Detail

Experiments performed 19 experiments Detail

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NMR combined restraints 4 contents Detail