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BMRB: 25755

2N69

NMR structure of non-sweet mutant (ins18RI19) of sweet protein Brazzein

Authors

Singarapu, K.K., Tonelli, M., Markley, J.L., Assadi-Porter, F.

Assembly

non-sweet mutant (ins18RI19) of sweet protein Brazzein

Entity

1. non-sweet mutant (ins18RI19) of sweet protein Brazzein (polymer), 56 monomers, 6767.618 Da Detail

QDKCKKVYEN YPVSKCQLRI ANQCNYDCKL DKHARSGECF YDEKRNLQCI CDYCEY

Formula weight

6767.618 Da

Source organism

Pentadiplandra brazzeana

Exptl. method

Refine. method

DGSA-distance geometry simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 100.0 %, Completeness: 79.1 %, Completeness (bb): 79.6 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	79.1 % (536 of 678)	88.4 % (319 of 361)	62.2 % (158 of 254)	93.7 % (59 of 63)
Backbone	79.6 % (266 of 334)	94.6 % (106 of 112)	64.7 % (108 of 167)	94.5 % (52 of 55)
Sidechain	80.7 % (322 of 399)	85.5 % (213 of 249)	71.8 % (102 of 142)	87.5 % (7 of 8)
Aromatic	35.5 % (22 of 62)	71.0 % (22 of 31)	0.0 % (0 of 31)
Methyl	100.0 % (32 of 32)	100.0 % (16 of 16)	100.0 % (16 of 16)

1. Brz RI

QDKCKKVYEN YPVSKCQLRI ANQCNYDCKL DKHARSGECF YDEKRNLQCI CDYCEY

Show sample condition

Sample

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

LACS Plot; CA

Referencing offset: -0.12 ppm, Outliers: 3 Detail

LACS Plot; CB

Referencing offset: -0.12 ppm, Outliers: 3 Detail

LACS Plot; HA

Referencing offset: -0.01 ppm, Outliers: 1 Detail

Protein Blocks Logo

Calculated from 20 models in PDB: 2N69, Strand ID: A Detail

Release date

2016-08-14

Related entities 1. non-sweet mutant (ins18RI19) of sweet protein Brazzein, : 1 : 19 entities Detail

Experiments performed 10 experiments Detail

1 2D 1H-15N HSQC

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

2 2D 1H-13C HSQC

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

3 3D CBCA(CO)NH

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

4 3D HNCO

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

5 3D HNCACB

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

6 3D HCCH-TOCSY

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

7 3D H(CCO)NH

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

8 3D 1H-15N NOESY

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

9 3D 1H-13C NOESY

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	D2O	natural abundance		10 %

10 3D 1H-13C NOESY aromatic

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 310 K, pH 5.2

#	Name	Isotope labeling	Type	Concentration
1	Brz_RI	[U-100% 13C; U-100% 15N]		1.5 mM
2	H2O	natural abundance		90 %
3	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_25755_2n69.nef
Input source #2: Coordindates	2n69.cif
Diamagnetism of the molecular assembly	True (excluding Oxygen atoms)
Whether the assembly has a disulfide bond	True (see coordinates for details)
Whether the assembly has a other bond	None
Whether the assembly contains a cyclic polymer	None
Overall data processing status	Warning

Disulfide bonds

Ptnr_site_1	Ptnr_site_2	Redox_state_prediction_1	Redox_state_prediction_2	Distance (Å)
A:4:CYS:SG	A:54:CYS:SG	oxidized, CA 57.415, CB 41.843 ppm	oxidized, CA 55.143, CB 44.382 ppm	1.95
A:16:CYS:SG	A:39:CYS:SG	oxidized, CA 56.57, CB 41.437 ppm	oxidized, CA 56.728, CB 45.019 ppm	2.114
A:24:CYS:SG	A:49:CYS:SG	oxidized, CA 58.406, CB 38.776 ppm	oxidized, CA 54.955, CB 38.445 ppm	1.983
A:28:CYS:SG	A:51:CYS:SG	oxidized, CA 60.914, CB 36.945 ppm	oxidized, CA 51.818, CB 35.982 ppm	1.956

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------
QDKCKKVYENYPVSKCQLRIANQCNYDCKLDKHARSGECFYDEKRNLQCICDYCEY
||||||||||||||||||||||||||||||||||||||||||||||||||||||||
QDKCKKVYENYPVSKCQLRIANQCNYDCKLDKHARSGECFYDEKRNLQCICDYCEY

Chain assignments

Entity_assembly_ID (NMR)	Auth_asym_ID (model)	Length	Unmapped	Conflict	Sequence coverage (%)
A	A	56	0	0	100.0

Content subtype: combined_25755_2n69.nef

#	Content subtype	Saveframe	Status	# of rows (sets)	Experiment type	Sequence coverage (%)
1	Covalent bonds	assembly	OK	4		No information
1	Assigned chemical shifts	assigned_chem_shift_list_1	Warning	545		100.0 (chain: A, length: 56)
1	Distance restraints	DYANA/DIANA_distance_constraints_2	Warning	1347 (1)	noe	96.4 (chain: A, length: 56)
1	Dihedral angle restraints	DYANA/DIANA_dihedral_3	OK	70 (70)	.	85.7 (chain: A, length: 56)

Assigned chemical shifts

Saveframe: assigned_chem_shift_list_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 56, Sequence coverage: 100.0%
- Total number of assignments
  - ¹H chemical shifts: 327
  - ¹³C chemical shifts: 159
  - ¹⁵N chemical shifts: 59
- Completeness of assignments
  - Entity_assemble_ID: A
- 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

Covalent bonds

Saveframe: assembly
- Status: OK

Distance restraints

Saveframe: DYANA/DIANA_distance_constraints_2
- Status: Warning
- Experiment type: noe
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 56, Sequence coverage: 96.4%
- Total number of restraints: 1347
- Weight of restraints: 1.0 (1347)
- Potential type of restraints: upper-bound-parabolic (1347)
- Range of distance target values: 2.27, 5.5 (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: DYANA/DIANA_dihedral_3
- Status: OK
- Experiment type: .
- Sequence coverage of experimental data
  - Entity_assembly_ID: A, Chain length: 56, Sequence coverage: 85.7%
- Total number of restraints: 70
- Total number of restraints per polymer type: 70
- Weight of restraints: 1.0 (70)
- Potential type of restraints: square-well-parabolic (70)
- Plot of Phi-Psi angle restraints
- Dihedral angle restraints per residue
  - Chain_ID: A

Keywords Brazzein, Sweet protein, non-sweet mutant ins18RI19