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BMRB: 6390

2AGM

NMR assignment of the R-module from the Azotobacter vinelandii mannuronan C5-epimerase AlgE4

Authors

Aachmann, F.L., Svanem, B.GLARUM., Valla, S., Petersen, S.BJORN., Wimmer, R.

Assembly

R-module

Entity

1. R-module (polymer, Thiol state: not present), 167 monomers, 16963.90 Da Detail

GSDGEPLVGG DTDDQLQGGS GADRLDGGAG DDILDGGAGR DRLSGGAGAD TFVFSAREDS YRTDTAVFND LILDFEASED RIDLSALGFS GLGDGYGGTL LLKTNAEGTR TYLKSFEADA EGRRFEVALD GDHTGDLSAA NVVFAATGTT TELEVLGDSG TQAGAIV

2. CA (non-polymer), 40.078 Da

Total weight

17003.979 Da

Max. entity weight

16963.9 Da

Source organism

Azotobacter vinelandii

Exptl. method

NMR

Refine. method

DISTANCE GEOMETRY, torsion angle dynamics, energy-minimisation

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 100.0 %, Completeness: 87.4 %, Completeness (bb): 96.0 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	87.4 % (1466 of 1677)	87.7 % (741 of 845)	85.5 % (564 of 660)	93.6 % (161 of 172)
Backbone	96.0 % (960 of 1000)	96.1 % (349 of 363)	96.8 % (456 of 471)	93.4 % (155 of 166)
Sidechain	78.6 % (640 of 814)	81.3 % (392 of 482)	74.2 % (242 of 326)	100.0 % (6 of 6)
Aromatic	22.2 % (24 of 108)	44.4 % (24 of 54)	0.0 % (0 of 54)
Methyl	93.5 % (174 of 186)	94.6 % (88 of 93)	92.5 % (86 of 93)

1. R-module

GSDGEPLVGG DTDDQLQGGS GADRLDGGAG DDILDGGAGR DRLSGGAGAD TFVFSAREDS YRTDTAVFND LILDFEASED RIDLSALGFS GLGDGYGGTL LLKTNAEGTR TYLKSFEADA EGRRFEVALD GDHTGDLSAA NVVFAATGTT TELEVLGDSG TQAGAIV

Show sample condition

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

LACS Plot; CA

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

Referencing offset: 0.27 ppm, Outliers: 1 Detail

LACS Plot; CB

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

Referencing offset: 0.27 ppm, Outliers: 1 Detail

LACS Plot; HA

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

Referencing offset: -0.11 ppm, Outliers: 2 Detail

LACS Plot; CO

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

Referencing offset: 1.06 ppm, Outliers: 5 Detail

Protein Blocks Logo

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

Release date

2005-04-07

Citation

NMR assignment of the R-module from the Azotobacter vinelandii Mannuronan C5-epimerase AlgE4
Aachmann, F.L., Svanem, B.GLAERUM., Valla, S., Petersen, S.BJORN., Wimmer, R.
J. Biomol. NMR (2005), 31, 259-259, PubMed 15803398 , DOI 10.1007/s10858-004-7911-7 , Abstract

Related entities 1. R-module, : 1 : 1 : 26 entities Detail

Experiments performed 5 experiments Detail

1 Aromatic side-chain atoms were assigned from a series of 2D H(CC)H-COSY, TOCSY

Instrument

Model Bruker DRX (600 MHz) Details Bruker DRX600 spectrometer equipped with a 5-mm xyz-grad TXI(H/C/N) probe.

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

2 and NOESY experiments optimized for aromatic signals.

Instrument

Model Bruker DRX (600 MHz) Details Bruker DRX600 spectrometer equipped with a 5-mm xyz-grad TXI(H/C/N) probe.

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

3 Due to the high content of Glycine and Alanine residues, MUSIC spectra have

Instrument

Model Bruker DRX (600 MHz) Details Bruker DRX600 spectrometer equipped with a 5-mm xyz-grad TXI(H/C/N) probe.

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

4 been very helpful to identify Gly/Ala residues and residues following after

Instrument

Model Bruker DRX (600 MHz) Details Bruker DRX600 spectrometer equipped with a 5-mm xyz-grad TXI(H/C/N) probe.

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

5 Gly/Ala.

Instrument

Model Bruker DRX (600 MHz) Details Bruker DRX600 spectrometer equipped with a 5-mm xyz-grad TXI(H/C/N) probe.

Sample #1

Temperature 298 (±0.2) K, pH 6.9 (±0.1), Details 95 % H2O, 5% D2O

#	Name	Isotope labeling	Concentration
1	R-module	[U-99% 13C; U-99% 15N]	1.6 mM
2	CALCIUM (II) ION		1.6 mM
3	CaCl2		50 mM
4	Na-HEPES		20 mM

Sample #2

Temperature 298 (±0.2) K, pH 6.9 (±0.1)

#	Name	Isotope labeling	Concentration
5	R-module	[U-99% 13C; U-99% 15N]	0.8 ~ 1.1 mM
6	CALCIUM (II) ION		0.8 ~ 1.1 mM
7	CaCl2		50 mM
8	Na-HEPES		20 mM

nullKeywords Alginate, Azotobacter vinelandii, R-module, Mannuronan C5-epimerase

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Found 1 Document (1.581 seconds)

BMRB: 6390

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Chemical shift reference

Chemical shift reference

Related entities 1. R-module, : 1 : 1 : 26 entities Detail

Experiments performed 5 experiments Detail

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