BMRBj - BMREntryMaster

Found 1 Document (0.467 seconds)

BMRB: 11428

6FRV

1H, 15N chemical shift assignments for a disulfide-deficient mutant of the starch-binding domain of glucoamylase

Authors

Sugimoto, H., Noda, Y., Segawa, S.

Assembly

a disulfide-deficient mutant of the starch-binding domain of glucoamylase

Entity

1. a disulfide-deficient mutant of the starch-binding domain of glucoamylase (polymer, Thiol state: not present), 110 monomers, 11975.77 Da Detail

TSGTTPTAVA VTFDLTATTT YGENIYLVGS ISQLGDWETS DGIALSADKY TSSDPLWYVT VTLPAGESFE YKFIRIESDD SVEWESDPNR EYTVPQAGGT STATVTDTWR

Formula weight

11975.77 Da

Source organism

Aspergillus niger

Exptl. method

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 96.4 %, Completeness: 85.0 %, Completeness (bb): 94.5 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹⁵N
All	85.0 % (604 of 711)	82.8 % (495 of 598)	96.5 % (109 of 113)
Backbone	94.5 % (310 of 328)	93.7 % (209 of 223)	96.2 % (101 of 105)
Sidechain	76.8 % (294 of 383)	76.3 % (286 of 375)	100.0 % (8 of 8)
Aromatic	95.5 % (64 of 67)	95.2 % (60 of 63)	100.0 % (4 of 4)
Methyl	95.4 % (62 of 65)	95.4 % (62 of 65)

1. a disulfide-deficient mutant of the starch-binding domain of glucoamylase

TSGTTPTAVA VTFDLTATTT YGENIYLVGS ISQLGDWETS DGIALSADKY TSSDPLWYVT VTLPAGESFE YKFIRIESDD SVEWESDPNR EYTVPQAGGT STATVTDTWR

Show sample condition

Sample

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

Release date

2011-09-27

Citation

NMR analysis of a kinetically trapped intermediate of a disulfide-deficient mutant of the starch-binding domain of glucoamylase
Sugimoto, H., Noda, Y., Segawa, S.
J. Mol. Biol. (2011), 412, 304-315, PubMed 21801731 , DOI 10.1016/j.jmb.2011.07.025 , Abstract

Related entities 1. a disulfide-deficient mutant of the starch-binding domain of glucoamylase, : 1 : 6 : 116 entities Detail

Experiments performed 6 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

2 3D 1H-15N NOESY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

3 3D 1H-15N TOCSY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

4 2D DQF-COSY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

5 3D 1H-15N HSQC-NOESY-HSQC

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

6 2D 1H-1H NOESY

Instrument

Bruker DRX - 600 MHz

Sample

State isotropic, Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 278 K, pH 7.0

#	Name	Isotope labeling	Type	Concentration
1	a disulfide-deficient mutant of the starch-binding domain of glucoamylase	[U-99% 15N]	protein	1 mM
2	sodium phosphate	natural abundance	buffer	20 mM
3	H2O		solvent	95 %
4	D2O		solvent	5 %

Chemical shift validation 3 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr11428_3.str
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

Content subtype: bmr11428_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chem_shift_list_1	Warning	725		96.4 (chain: 1, length: 110)
1	Chemical shift references	chemical_shift_reference_1	OK	2		No information

Assigned chemical shifts

Saveframe: assigned_chem_shift_list_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 110, Sequence coverage: 96.4%
- Total number of assignments
  - ¹H chemical shifts: 616
  - ¹⁵N chemical shifts: 109
- Completeness of assignments
  - Entity_assemble_ID: 1
- Histogram of normalized assigned chemical shifts
- Random coil index and derived parameters (S² and NMR RMSD)
  - Chain_ID: 1

Keywords aromatic cluster, disulfide bond, NMR, refolding intermediate, starch-binding domain