SASDAH6: WbdD(1-459) (bifunctional kinase- methyltransferase WbdD, WbdD)

Basic information

• Entry: Database: SASBDB / ID: SASDAH6

Sample

WbdD(1-459)

• bifunctional kinase- methyltransferase WbdD (protein), WbdD, Escherichia coli

• Biological species: Escherichia coli (E. coli)
• Citation: Journal: Nat Struct Mol Biol / Year: 2015; Title: A coiled-coil domain acts as a molecular ruler to regulate O-antigen chain length in lipopolysaccharide.; Authors: Gregor Hagelueken / Bradley R Clarke / Hexian Huang / Anne Tuukkanen / Iulia Danciu / Dmitri I Svergun / Rohanah Hussain / Huanting Liu / Chris Whitfield / James H Naismith / ; Abstract: Long-chain bacterial polysaccharides have important roles in pathogenicity. In Escherichia coli O9a, a model for ABC transporter-dependent polysaccharide assembly, a large extracellular carbohydrate with a narrow size distribution is polymerized from monosaccharides by a complex of two proteins, WbdA (polymerase) and WbdD (terminating protein). Combining crystallography and small-angle X-ray scattering, we found that the C-terminal domain of WbdD contains an extended coiled-coil that physically separates WbdA from the catalytic domain of WbdD. The effects of insertions and deletions in the coiled-coil region were analyzed in vivo, revealing that polymer size is controlled by varying the length of the coiled-coil domain. Thus, the coiled-coil domain of WbdD functions as a molecular ruler that, along with WbdA:WbdD stoichiometry, controls the chain length of a model bacterial polysaccharide.

Contact author

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

Models

• Model #189: ; Type: mix / Software: CORAL / Radius of dummy atoms: 1.90 A; Search similar-shape structures of this assembly by Omokage search (details)
• Model #192: ; Type: dummy / Software: DAMMIN / Radius of dummy atoms: 1.90 A; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: WbdD(1-459) / Contrast: 3.047 / Dry vol: 59200 / Specimen concentration: 1.00-10.00 / Concentration method: Nanodrop
• Buffer: Name: BisTris / Concentration: 20.00 mM / pH: 7 / Composition: 50 mM NaCl, 5 mM DTT
• Entity #126: Name: WbdD / Type: protein / Description: bifunctional kinase- methyltransferase WbdD / Formula weight: 59.2 / Num. of mol.: 1 / Source: Escherichia coli

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Shape: 0.6 / Type of source: X-ray synchrotron / Wavelength: 0.15 Å / Dist. spec. to detc.: 2.7 mm
• Detector: Name: Pilatus 1M-W / Pixsize x: 0.172 mm

Scan

Title: WbdD(1-459) / Measurement date: Sep 23, 2011 / Storage temperature: 20 °C / Cell temperature: 20 °C / Exposure time: 15 sec. / Number of frames: 10 / Unit: 1/nm /

	Min	Max
Q	0.0855	6.0292

Distance distribution function P(R)

Sofotware P(R): GNOM 4.5a / Number of points: 435 /

	Min	Max
Q	0.1828	2.563
P(R) point	36	470
R	0	9.979

Result

D max: 10 / Type of curve: merged / Standard: BSA /

	Experimental	Standard	Porod
MW	52 kDa	52 kDa	53 kDa
Volume	-	-	90 nm3

	P(R)	Guinier
Forward scattering, I0	47.5	47.4
Radius of gyration, Rg	3.2 nm	3.1 nm