BMRBj - BMREntryMaster

Found 1 Document (0.673 seconds)

BMRB: 30765

6XFJ

Structural characterization of the type III secretion system pilotin-secretin complex InvH-InvG by NMR spectroscopy

Authors

Majewski, D.D., Okon, M., Heinkel, F., Robb, C.S., Vuckovic, M., McIntosh, L.P., Strynadka, N.C.J.

Assembly

Type 3 secretion system pilotin, Type 3 secretion system secretin

Entity

1. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 1 (polymer, Thiol state: all free), 82 monomers, 9440.546 Da Detail

GSHMDNSASK NSAISSSIFC EKYKQTKEQA LTFFQEHPQY MRSKEDEEQL MTEFKKVLLE PGSKNLSIYQ TLLAAHERLQ AL

2. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 2 (polymer, Thiol state: not present), 47 monomers, 5241.760 Da Detail

GSHMDPLTPD ASESVNNILK QSGAWSGDDK LQKWVRVYLD RGQEAIK

Total weight

14682.306 Da

Max. entity weight

9440.546 Da

Source organism

Salmonella enterica subsp. enterica serovar Typhimurium str. LT2

Exptl. method

solution NMR

Data set

assigned_chemical_shifts, spectral_peak_list

Chem. Shift Complete

Sequence coverage: 96.1 %, Completeness: 87.1 %, Completeness (bb): 87.2 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	87.1 % (1335 of 1532)	83.2 % (668 of 803)	91.8 % (535 of 583)	90.4 % (132 of 146)
Backbone	87.2 % (668 of 766)	80.4 % (209 of 260)	90.0 % (343 of 381)	92.8 % (116 of 125)
Sidechain	87.4 % (777 of 889)	83.4 % (453 of 543)	94.8 % (308 of 325)	76.2 % (16 of 21)
Aromatic	96.4 % (108 of 112)	96.4 % (54 of 56)	96.3 % (52 of 54)	100.0 % (2 of 2)
Methyl	95.7 % (111 of 116)	91.4 % (53 of 58)	100.0 % (58 of 58)

1. entity 1

GSHMDNSASK NSAISSSIFC EKYKQTKEQA LTFFQEHPQY MRSKEDEEQL MTEFKKVLLE PGSKNLSIYQ TLLAAHERLQ AL

2. entity 2

GSHMDPLTPD ASESVNNILK QSGAWSGDDK LQKWVRVYLD RGQEAIK

Show sample condition

Sample #1

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 20 mM NA- MOPS, 150 mM NA- sodium chloride, 0.2 mM NA- TCEP, 10 % v/v NA- D2O, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

Sample #2

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 20 mM NA- MOPS, 150 mM NA- sodium chloride, 0.2 mM NA- TCEP, 10 % v/v NA- D2O, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

Release date

2020-06-15

Citation

Characterization of the Pilotin-Secretin Complex from the Salmonella enterica Type III Secretion System Using Hybrid Structural Methods
Majewski, D.D., Okon, M., Heinkel, F., Robb, C.S., Vuckovic, M., McIntosh, L.P., Strynadka, N.C.J.
Structure (2020), 29, 125-138, PubMed 32877645 , DOI 10.1016/j.str.2020.08.006 , Abstract

Related entities 1. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 1, : 1 : 1 : 3 entities Detail

Related entities 2. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 2, : 2 entities Detail

Interaction partners 2. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 2, : 3 interactors Detail

More details for 3 interactors identified by 1 citations

Experiments performed 33 experiments Detail

1 2D 1H-15N HSQC

Instrument

Bruker AVANCE - 500 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 20 mM NA- MOPS, 150 mM NA- sodium chloride, 0.2 mM NA- TCEP, 10 % v/v NA- D2O, 90% H2O/10% D2O

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

2 3D HNCACB

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

3 3D CBCA(CO)NH

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

4 3D HN(CA)CO

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

5 3D HCCH-TOCSY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

6 3D HNCO

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

7 3D HCC(CO)NH-TOCSY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

8 3D hbCBcgcCH

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

9 3D hbCBcgcdceHE

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

10 3D hbCBcgcdHD

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

11 3D HBHA(CBCACO)NH

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

12 2D 1H-13C HSQC

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

13 3D 1H-15N NOESY

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

14 3D 1H-13C NOESY aromatic

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

15 3D HNCACB

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

16 3D HNCO

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

17 2D 1H-15N HSQC

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

18 3D 1H-15N NOESY filtered-edited

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

19 3D 1H-13C NOESY filtered-edited

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

20 3D 1H-15N NOESY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

21 3D 1H-13C NOESY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
1	entity_1	[U-13C; U-15N]	0.5 mM
2	entity_2	[U-13C; U-15N]	0.5 mM
3	MOPS	natural abundance	20 mM
4	sodium chloride	natural abundance	150 mM
5	TCEP	natural abundance	0.2 mM
6	D2O	natural abundance	10 % v/v

22 3D 1H-13C NOESY aliphatic

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

23 3D hbCBcgcCH

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

24 3D hbCBcgcdceHE

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

25 3D hbCBcgcdHD

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

26 3D HBHA(CBCACO)NH

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

27 2D 1H-13C HSQC

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

28 3D HCC(CO)NH-TOCSY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

29 3D HCCH-TOCSY

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

30 3D HN(CA)CO

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

31 3D CBCA(CO)NH

Instrument

Bruker AVANCE - 850 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

32 3D 1H-15N NOESY filtered-edited

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

33 3D 1H-13C NOESY filtered-edited

Instrument

Bruker AVANCE - 500 MHz

Sample

#	Name	Isotope labeling	Concentration
7	entity_1	[U-13C; U-15N]	0.5 mM
8	entity_2	[U-13C; U-15N]	0.5 mM
9	MOPS	natural abundance	20 mM
10	sodium chloride	natural abundance	150 mM
11	TCEP	natural abundance	0.2 mM
12	D2O	natural abundance	10 % v/v

Chemical shift validation 4 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr30765_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: bmr30765_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chemical_shifts_1	Warning	1561		96.3 (chain: 1, length: 82), 95.7 (chain: 2, length: 47)
1	Chemical shift references	chem_shift_reference_1	OK	3		No information

Assigned chemical shifts

Saveframe: assigned_chemical_shifts_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 82, Sequence coverage: 96.3%
  - Entity_assembly_ID: 2, Chain length: 47, Sequence coverage: 95.7%
- Total number of assignments
  - ¹H chemical shifts: 876
  - ¹³C chemical shifts: 551
  - ¹⁵N chemical shifts: 134
- Completeness of assignments
  - Entity_assemble_ID: 1
  - Entity_assemble_ID: 2
- Redox state of CYS
- Peptide bond of PRO
- Tautomeric state of HIS
- 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
  - Chain_ID: 2

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	520	498	95.8
¹³C chemical shifts	378	361	95.5
¹⁵N chemical shifts	92	85	92.4

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	164	156	95.1
¹³C chemical shifts	164	154	93.9
¹⁵N chemical shifts	80	76	95.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	356	342	96.1
¹³C chemical shifts	214	207	96.7
¹⁵N chemical shifts	12	9	75.0

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	39	36	92.3
¹³C chemical shifts	39	36	92.3

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	38	36	94.7
¹³C chemical shifts	38	36	94.7

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	283	262	92.6
¹³C chemical shifts	205	190	92.7
¹⁵N chemical shifts	54	49	90.7

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	96	88	91.7
¹³C chemical shifts	94	85	90.4
¹⁵N chemical shifts	45	42	93.3

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	187	174	93.0
¹³C chemical shifts	111	105	94.6
¹⁵N chemical shifts	9	7	77.8

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	23	22	95.7
¹³C chemical shifts	23	22	95.7

Atom group	# of target shifts	# of assigned shifts	Completeness (%)
¹H chemical shifts	18	18	100.0
¹³C chemical shifts	16	16	100.0
¹⁵N chemical shifts	2	2	100.0

Keywords PROTEIN TRANSPORT, chaperone, pilotin, secretin, type III secretion

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Found 1 Document (0.673 seconds)

BMRB: 30765

Sample #1

Sample #2

Related entities 1. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 1, : 1 : 1 : 3 entities Detail

Related entities 2. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 2, : 2 entities Detail

Interaction partners 2. Type 3 secretion system pilotin, Type 3 secretion system secretin, entity 2, : 3 interactors Detail

Experiments performed 33 experiments Detail

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