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BMRB: 19367

2MAP

Solution structure of the complex formed by the region 2 of E. coli sigmaE and its cognate -10 non template element TGTCAAA

Authors

Campagne, S., Vorholt, J.A., Allain, F.H-T.

Assembly

complex formed by the region 2 of E. coli sigmaE and its cognate -10 non template element TGTCAAA

Entity

1. complex formed by the region 2 of E. coli sigmaE and its cognate -10 non template element TGTCAAA, entity 1 (polymer, Thiol state: not present), 96 monomers, 11010.47 Da Detail

MSEQLTDQVL VERVQKGDQK AFNLLVVRYQ HKVASLVSRY VPSGDVPDVV QEAFIKAYRA LDSFRGDSAF YTWLYRIAVN TAKNYLVAQG RRLELV

2. complex formed by the region 2 of E. coli sigmaE and its cognate -10 non template element TGTCAAA, entity 2 (polymer), 7 monomers, 2184.408 Da Detail

TGTCAAA

Total weight

13194.878 Da

Max. entity weight

11010.47 Da

Source organism

Escherichia coli

Exptl. method

Refine. method

simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 93.2 %, Completeness: 82.2 %, Completeness (bb): 76.5 % Detail

Polymer type: polypeptide(L) polydeoxyribonucleotide

	Total	¹H	¹³C	¹⁵N
All	82.2 % (1000 of 1216)	86.4 % (571 of 661)	72.4 % (326 of 450)	98.1 % (103 of 105)
Backbone	76.5 % (475 of 621)	79.8 % (194 of 243)	66.5 % (189 of 284)	97.9 % (92 of 94)
Sidechain	89.4 % (614 of 687)	90.4 % (378 of 418)	87.2 % (225 of 258)	100.0 % (11 of 11)
Aromatic	72.9 % (86 of 118)	81.8 % (54 of 66)	60.8 % (31 of 51)	100.0 % (1 of 1)
Methyl	93.1 % (121 of 130)	90.9 % (60 of 66)	95.3 % (61 of 64)

1. entity 1

MSEQLTDQVL VERVQKGDQK AFNLLVVRYQ HKVASLVSRY VPSGDVPDVV QEAFIKAYRA LDSFRGDSAF YTWLYRIAVN TAKNYLVAQG RRLELV

2. entity 2

TGTCAAA

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

Sample #2

Solvent system 100% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
7	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
8	entity_2	natural abundance		0.8 mM
9	D2O	natural abundance		100 %
10	NaPO4	natural abundance		10 mM
11	NaCl	natural abundance		50 mM

LACS Plot; CA

Referencing offset: 0.05 ppm, Outliers: 2 Detail

LACS Plot; CB

Referencing offset: 0.05 ppm, Outliers: 2 Detail

LACS Plot; HA

Referencing offset: 0.02 ppm, Outliers: 2 Detail

Protein Blocks Logo

Calculated from 18 models in PDB: 2MAP, Strand ID: A Detail

Release date

2014-02-09

Citation

Structural basis for -10 promoter element melting by environmentally induced sigma factors
Campagne, S., Marsh, M.E., Capitani, G., Vorholt, J.A., Allain, F.
Nat. Struct. Mol. Biol. (2014), 21, 269-276, PubMed 24531660 , DOI 10.1038/nsmb.2777 , Abstract

Related entities 1. complex formed by the region 2 of E. coli sigmaE and its cognate -10 non template element TGTCAAA, entity 1, : 1 : 2 : 3 : 14 entities Detail

Experiments performed 14 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

2 2D 1H-13C HSQC

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

3 2D 1H-13C HSQC aromatic

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

4 3D HNCACB

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

5 3D CBCA(CO)NH

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

6 3D HNCO

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

7 3D HNCA

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

8 3D HN(CO)CA

Instrument

Bruker Avance - 500 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

9 3D HCCH-TOCSY

Instrument

Bruker Avance - 700 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

10 3D 1H-15N NOESY

Instrument

Bruker Avance - 900 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

11 3D 1H-13C NOESY aliphatic

Instrument

Bruker Avance - 900 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

12 3D 1H-13C NOESY aromatic

Instrument

Bruker Avance - 900 MHz cryo probbed

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
1	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
2	entity_2	natural abundance		0.8 mM
3	H2O	natural abundance		90 %
4	D2O	natural abundance		10 %
5	NaPO4	natural abundance		10 mM
6	NaCl	natural abundance		50 mM

13 2D NOESY 1H-1H F1F2F

Instrument

Bruker Avance - 700 MHz cryo probbed

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
7	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
8	entity_2	natural abundance		0.8 mM
9	D2O	natural abundance		100 %
10	NaPO4	natural abundance		10 mM
11	NaCl	natural abundance		50 mM

14 2D NOESY 1H-1H F2F

Instrument

Bruker Avance - 700 MHz cryo probbed

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 303 K, pH 6.5

#	Name	Isotope labeling	Type	Concentration
7	entity_1	[U-100% 13C; U-100% 15N]		0.8 mM
8	entity_2	natural abundance		0.8 mM
9	D2O	natural abundance		100 %
10	NaPO4	natural abundance		10 mM
11	NaCl	natural abundance		50 mM

nullKeywords -10 element recognition, ECF sigma factor, Protein-ssDNA complex