SASDC98: Surface presentation of antigens protein SpaOc-SpaO N-terminus

Basic information

Entry

Database: SASBDB / ID: SASDC98

Sample

Surface presentation of antigens protein SpaOc-SpaO N-terminus

• Surface presentation of antigens protein SpaO SpaO(SPOA2) (protein), SpaOc, Salmonella enterica subsp. enterica serovar Typhimurium
• Surface presentation of antigens protein SpaO(SPOA1,2) N-terminus (protein), SpaO N-terminus, Salmonella enterica subsp. enterica serovar Typhimurium

• Function / homology: Type III secretion system apparatus protein YscQ/HrcQ/SpaO / Type III secretion system outer membrane, SpaO / Flagellar motor switch protein FliN-like, C-terminal domain / SpoA-like superfamily / Type III flagellar switch regulator (C-ring) FliN C-term / bacterial-type flagellum-dependent swarming motility / protein secretion by the type III secretion system / positive chemotaxis / Surface presentation of antigens protein SpaO
• Biological species: Salmonella enterica subsp. enterica serovar Typhimurium (bacteria)
• Citation: Journal: J Mol Biol / Year: 2019; Title: Molecular Organization of Soluble Type III Secretion System Sorting Platform Complexes.; Authors: Ivonne Bernal / Jonathan Börnicke / Johannes Heidemann / Dmitri Svergun / Julia A Horstmann / Marc Erhardt / Anne Tuukkanen / Charlotte Uetrecht / Michael Kolbe / ; Abstract: Many medically relevant Gram-negative bacteria use the type III secretion system (T3SS) to translocate effector proteins into the host for their invasion and intracellular survival. A multi-protein complex located at the cytosolic interface of the T3SS is proposed to act as a sorting platform by selecting and targeting substrates for secretion through the system. However, the precise stoichiometry and 3D organization of the sorting platform components are unknown. Here we reconstitute soluble complexes of the Salmonella Typhimurium sorting platform proteins including the ATPase InvC, the regulator OrgB, the protein SpaO and a recently identified subunit SpaO, which we show to be essential for the solubility of SpaO. We establish domain-domain interactions, determine for the first time the stoichiometry of each subunit within the complexes by native mass spectrometry and gain insight into their organization using small-angle X-ray scattering. Importantly, we find that in solution the assembly of SpaO/SpaO/OrgB/InvC adopts an extended L-shaped conformation resembling the sorting platform pods seen in in situ cryo-electron tomography, proposing that this complex is the core building block that can be conceivably assembled into higher oligomers to form the T3SS sorting platform. The determined molecular arrangements of the soluble complexes of the sorting platform provide important insights into its architecture and assembly.

Contact author

• Anne Tuukkanen (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

Models

Sample

• Sample: Name: Surface presentation of antigens protein SpaOc-SpaO N-terminus; Specimen concentration: 9.5 mg/ml / Entity id: 824 / 826
• Buffer: Name: 20 mM HEPES pH 7.5, 150 mM NaCl / pH: 7.5

Entity #824

Name: SpaOc / Type: protein
Description: Surface presentation of antigens protein SpaO SpaO(SPOA2)
Formula weight: 12.374 / Num. of mol.: 2
Source: Salmonella enterica subsp. enterica serovar Typhimurium
References: UniProt: P40699
Sequence:

METLDIQHIE EENNTTETAE TLPGLNQLPV KLEFVLYRKN VTLAELEAMG QQQLLSLPTN AELNVEIMAN GVLLGNGELV QMNDTLGVEI HEWLSESGNG ESAWSHPQFE K

Entity #826

Name: SpaO N-terminus / Type: protein
Description: Surface presentation of antigens protein SpaO(SPOA1,2) N-terminus
Formula weight: 17.423 / Num. of mol.: 1
Source: Salmonella enterica subsp. enterica serovar Typhimurium
References: UniProt: P40699
Sequence:

MSLRVRQIDR REWLLAQTAT ECQRHGREAT LEYPTRQGMW VRLSDAEKRW SAWIKPGDWL EHVSPALAGA AVSAGAEHLV VPWLAATERP FELPVPHLSC RRLCVENPVP GSALPEGKLL HIMSDRGGLW FEHLPELPAV GGGRPSAWSH PQFEK

Experimental information

• Beam: Instrument name: PETRA III EMBL P12 / City: Hamburg / 国: Germany / Type of source: X-ray synchrotron / Wavelength: 0.124 Å
• Detector: Name: Pilatus 2M

Scan

Title: Surface presentation of antigens protein SpaOc-SpaO N-terminus
Measurement date: Apr 24, 2017 / Storage temperature: 20 °C / Cell temperature: 20 °C / Number of frames: 3600 / Unit: 1/nm /

	Min	Max
Q	0.1852	5.0254

Distance distribution function P(R)

Sofotware P(R): GNOM 5.0 / Number of points: 911 /

	Min	Max
Q	0.237428	2.73716
P(R) point	1	911
R	0	10.64

Result

Type of curve: single_conc /

	Experimental	Porod
MW	36.3 kDa	-
Volume	-	68 nm3

	P(R)	P(R) error	Guinier	Guinier error
Forward scattering, I0	4847	7.5	4759.4	11
Radius of gyration, Rg	3.06 nm	0.005	2.93 nm	0.008

	Min	Max
D	-	10.64
Guinier point	27	94