BMRBj - BMREntryMaster

Found 1 Document (1.259 seconds)

BMRB: 4354

7MN6

Solution NMR Studies of a 42kDa Escherichia coli Maltose Binding Protein/ Beta-Cyclodextrin Complex: Chemical Shift Assignments and Analysis

Authors

Gardner, K.H., Zhang, X., Gehring, K., Kay, L.E.

Assembly

E. coli maltose binding protein

Entity

1. E. coli maltose binding protein (polymer), 370 monomers, 40694.71 Da Detail

KTEEGKLVIW INGDKGYNGL AEVGKKFEKD TGIKVTVEHP DKLEEKFPQV AATGDGPDII FWAHDRFGGY AQSGLLAEIT PDKAFQDKLY PFTWDAVRYN GKLIAYPIAV EALSLIYNKD LLPNPPKTWE EIPALDKELK AKGKSALMFN LQEPYFTWPL IAADGGYAFK YENGKYDIKD VGVDNAGAKA GLTFLVDLIK NKHMNADTDY SIAEAAFNKG ETAMTINGPW AWSNIDTSKV NYGVTVLPTF KGQPSKPFVG VLSAGINAAS PNKELAKEFL ENYLLTDEGL EAVNKDKPLG AVALKSYEEE LAKDPRIAAT MENAQKGEIM PNIPQMSAFW YAVRTAVINA ASGRQTVDEA LKDAQTRITK

2. BCD (non-polymer), 1134.984 Da

Total weight

41829.695 Da

Max. entity weight

40694.71 Da

Source organism

Escherichia coli

Exptl. method

NMR

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 97.0 %, Completeness: 55.4 %, Completeness (bb): 81.6 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	55.4 % (2392 of 4316)	36.0 % (806 of 2237)	74.0 % (1252 of 1692)	86.3 % (334 of 387)
Backbone	81.6 % (1778 of 2178)	58.2 % (435 of 748)	93.7 % (1013 of 1081)	94.6 % (330 of 349)
Sidechain	38.0 % (941 of 2479)	24.9 % (371 of 1489)	59.5 % (566 of 952)	10.5 % (4 of 38)
Aromatic	5.6 % (21 of 378)	9.5 % (18 of 189)	1.7 % (3 of 181)	0.0 % (0 of 8)
Methyl	83.3 % (348 of 418)	77.5 % (162 of 209)	89.0 % (186 of 209)

1. E. coli maltose binding protein

Show sample condition

Sample

Temperature 310 (±0.2) K, pH 7.2 (±0.05), Details Exchangeable proton sites in MBP were fully protonated by partially unfolding this protein in 2.5M GuHCl at room temperature for 3hr and subsequently refolding in GuHCl-free phosphate buffer containing 2mM beta-cyclodextrin. The protein under study has high level (>95%) deuterium labeling at all aliphatic and aromatic sites except at the following methyl groups which are highly protonated: Ile d1, Val g1/g2 and Leu d1/d2. No attempt has been made here to correct the 2H isotope effects on the 15N and 13C chemical shifts that are reported.

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

Release date

1999-06-09

Citation

Solution NMR Studies of a 42kDa Escherichia coli Maltose Binding Protein/ Beta-Cyclodextrin Complex: Chemical Shift Assignments and Analysis
Gardner, K.H., Zhang, X., Gehring, K., Kay, L.E.
J. Am. Chem. Soc. (1998), 120, 11738-11748, PubMed , DOI 10.1021/ja982019w

Related entities 1. E. coli maltose binding protein, : 1 : 4 : 95 : 13 entities Detail

Interaction partners 1. E. coli maltose binding protein, : 1 interactors Detail

More details for 1 interactors identified by 1 citations

Experiments performed 11 experiments Detail

1 1H-15N HSQC

Instrument

Varian INOVA - 600 MHz

Sample

State isotropic, Temperature 310 (±0.2) K, pH 7.2 (±0.05), Details Exchangeable proton sites in MBP were fully protonated by partially unfolding this protein in 2.5M GuHCl at room temperature for 3hr and subsequently refolding in GuHCl-free phosphate buffer containing 2mM beta-cyclodextrin. The protein under study has high level (>95%) deuterium labeling at all aliphatic and aromatic sites except at the following methyl groups which are highly protonated: Ile d1, Val g1/g2 and Leu d1/d2. No attempt has been made here to correct the 2H isotope effects on the 15N and 13C chemical shifts that are reported.

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

2 CT-HNCA

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

3 CT-HN(CO)CA

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

4 CT-HN(CA)CB

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

5 CT-HN(COCA)CB

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

6 HNCO

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

7 CT-1H-13C HSQC

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

8 (H)C(CO)NH-TOCSY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

9 H(CCO)NH-TOCSY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

10 (H)C(CA)NH-TOCSY

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

11 (HM)CMC(CM)HM

Instrument

Varian INOVA - 600 MHz

Sample

#	Name	Isotope labeling	Concentration
1	E. coli maltose binding protein	[U-15N;U-13C; U-2H;1H-95% Ile HD1, 1H-50% LeuHD1/HD2, 1H-85% Val HG1/HG2]	0.7 ~ 0.9 mM
2	BCD		2.0 mM
3	sodium phosphate buffer		20 mM
4	EDTA		100 uM
5	Pefabloc		0.1 mg/mL
6	pepstatin		1.0 ug/uL
7	D2O		10 %
8	H2O		90 %

Chemical shift validation 4 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr4354_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: bmr4354_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	shift_set_1	Warning	2280		96.5 (chain: 1, length: 370)
1	Chemical shift references	chemical_shift_reference	Warning	3		No information

Assigned chemical shifts

Saveframe: shift_set_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 370, Sequence coverage: 96.5%
- Total number of assignments
  - ¹³C chemical shifts: 1198
  - ¹⁵N chemical shifts: 330
  - ¹H chemical shifts: 752
- Completeness of assignments
  - Entity_assemble_ID: 1
- 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: 1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	1692	1198	70.8
¹⁵N chemical shifts	393	330	84.0
¹H chemical shifts	2237	472	21.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	740	682	92.2
¹⁵N chemical shifts	349	330	94.6
¹H chemical shifts	748	330	44.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	952	516	54.2
¹⁵N chemical shifts	44	0	0.0
¹H chemical shifts	1489	142	9.5

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	215	183	85.1
¹H chemical shifts	215	140	65.1

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹³C chemical shifts	181	0	0.0
¹⁵N chemical shifts	8	0	0.0
¹H chemical shifts	189	2	1.1

Keywords deuteration, MBP

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Found 1 Document (1.259 seconds)

BMRB: 4354

Sample

Related entities 1. E. coli maltose binding protein, : 1 : 4 : 95 : 13 entities Detail

Interaction partners 1. E. coli maltose binding protein, : 1 interactors Detail

Experiments performed 11 experiments Detail

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Chemical shift validation 4 contents Detail