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BMRB: 19023

2M4V

Solution structure and sigma factor interactions of Mycobacterium tuberculosis RNA polymerase binding protein A

Authors

Bortoluzzi, A., Muskett, F.W., Waters, L.C., Adiss, P.W., Rieck, B., Munder, T., Schleier, S., Forti, F., Ghisotti, D., Carr, M.D., O'Hare, H.M.

Assembly

RNA polymerase binding protein A (RbpA)

Entity

1. RNA polymerase binding protein A (RbpA) (polymer, Thiol state: all free), 80 monomers, 9012.963 Da Detail

SMADRVLRGS RLGAVSYETD RNHDLAPRQI ARYRTDNGEE FEVPFADDAE IPGTWLCRNG MEGTLIEGDL PEPKKVKPPR

Formula weight

9012.963 Da

Source organism

Mycobacterium tuberculosis

Exptl. method

solution NMR

Refine. method

simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 96.2 %, Completeness: 83.4 %, Completeness (bb): 89.3 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	83.4 % (758 of 909)	87.9 % (422 of 480)	74.6 % (262 of 351)	94.9 % (74 of 78)
Backbone	89.3 % (416 of 466)	93.1 % (149 of 160)	85.0 % (198 of 233)	94.5 % (69 of 73)
Sidechain	78.9 % (407 of 516)	85.3 % (273 of 320)	67.5 % (129 of 191)	100.0 % (5 of 5)
Aromatic	46.2 % (24 of 52)	88.5 % (23 of 26)	0.0 % (0 of 25)	100.0 % (1 of 1)
Methyl	94.4 % (68 of 72)	100.0 % (36 of 36)	88.9 % (32 of 36)

1. entity

SMADRVLRGS RLGAVSYETD RNHDLAPRQI ARYRTDNGEE FEVPFADDAE IPGTWLCRNG MEGTLIEGDL PEPKKVKPPR

Show sample condition

Sample #1

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	RbpA	[U-15N]	0.3 ~ 0.35 mM
2	potassium chloride	natural abundance	100 mM
3	potassium phosphate	natural abundance	25 mM
4	DTT	natural abundance	0.5 mM
5	EDTA	natural abundance	0.5 mM
6	sodium azide	natural abundance	0.02 % w/v
7	TCEP	natural abundance	0.5 mM
8	AEBSF protease inhibitor	natural abundance	0.2 mM

Sample #2

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5

#	Name	Isotope labeling	Concentration
9	RbpA	[U-13C; U-15N]-His-Phe-Tyr-Trp	0.3 ~ 0.35 mM
10	potassium chloride	natural abundance	100 mM
11	potassium phosphate	natural abundance	25 mM
12	DTT	natural abundance	0.5 mM
13	EDTA	natural abundance	0.5 mM
14	sodium azide	natural abundance	0.02 % w/v
15	TCEP	natural abundance	0.5 mM
16	AEBSF protease inhibitor	natural abundance	0.2 mM

Sample #3

Solvent system 95% H2O/5% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5

#	Name	Isotope labeling	Concentration
17	RbpA	[U-15N]	0.3 ~ 0.35 mM
18	potassium chloride	natural abundance	100 mM
19	potassium phosphate	natural abundance	25 mM
20	DTT	natural abundance	0.5 mM
21	EDTA	natural abundance	0.5 mM
22	sodium azide	natural abundance	0.02 % w/v
23	TCEP	natural abundance	0.5 mM
24	AEBSF protease inhibitor	natural abundance	0.2 mM

LACS Plot; CA

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

Referencing offset: -0.11 ppm, Outliers: 1 Detail

LACS Plot; CB

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

Referencing offset: -0.11 ppm, Outliers: 1 Detail

LACS Plot; HA

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

Referencing offset: 0.01 ppm, Outliers: 2 Detail

LACS Plot; CO

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

Referencing offset: 0.43 ppm, Outliers: 2 Detail

Protein Blocks Logo

Calculated from 35 models in PDB: 2M4V, Strand ID: A Detail

Release date

2013-04-10

Citation

Mycobacterium tuberculosis RNA polymerase-binding protein A (RbpA) and its interactions with sigma factors
Bortoluzzi, A., Muskett, F.W., Waters, L.C., Adiss, P.W., Rieck, B., Munder, T., Schleier, S., Forti, F., Ghisotti, D., Carr, M.D., O'Hare, H.M.
J. Biol. Chem. (2013), 288, 14438-14450, PubMed 23548911 , DOI 10.1074/jbc.M113.459883 , Abstract

Entries sharing articles Swiss-Prot: 3 entries Detail

  Swiss-Prot: P9WGI1 released on 2014-04-16
    Title SIGA_MYCTU Entity RNA polymerase sigma factor SigA
  Swiss-Prot: P9WGI5 released on 2014-04-16
    Title SIGB_MYCTU Entity RNA polymerase sigma factor SigB
  Swiss-Prot: P9WHJ5 released on 2014-04-16
    Title RBPA_MYCTU Entity RNA polymerase-binding protein RbpA

Related entities 1. RNA polymerase binding protein A (RbpA), : 1 : 17 : 12 entities Detail

Experiments performed 8 experiments Detail

1 2D 1H-1H TOCSY

Instrument

Bruker Avance - 600 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	RbpA	[U-15N]	0.3 ~ 0.35 mM
2	potassium chloride	natural abundance	100 mM
3	potassium phosphate	natural abundance	25 mM
4	DTT	natural abundance	0.5 mM
5	EDTA	natural abundance	0.5 mM
6	sodium azide	natural abundance	0.02 % w/v
7	TCEP	natural abundance	0.5 mM
8	AEBSF protease inhibitor	natural abundance	0.2 mM

2 2D 1H-1H NOESY

Instrument

Bruker Avance - 600 MHz

Sample

State isotropic, Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5

#	Name	Isotope labeling	Concentration
1	RbpA	[U-15N]	0.3 ~ 0.35 mM
2	potassium chloride	natural abundance	100 mM
3	potassium phosphate	natural abundance	25 mM
4	DTT	natural abundance	0.5 mM
5	EDTA	natural abundance	0.5 mM
6	sodium azide	natural abundance	0.02 % w/v
7	TCEP	natural abundance	0.5 mM
8	AEBSF protease inhibitor	natural abundance	0.2 mM

3 2D 1H-15N HSQC