Characterization of the Pilotin-Secretin Complex from the Salmonella enterica Type III Secretion System Using Hybrid Structural Methods.

Majewski DD, Okon M, Heinkel F, Robb CS, Vuckovic M, McIntosh LP, Strynadka NCJ, Structure (2020) Europe PMC

SASDJS4 – Type 3 secretion system pilotin, InvH70-147

Type 3 secretion system pilotin
MWexperimental 18 kDa
MWexpected 19 kDa
VPorod 29 nm3
log I(s) 8.60×100 8.60×10-1 8.60×10-2 8.60×10-3
Type 3 secretion system pilotin small angle scattering data  s, nm-1
ln I(s)
Type 3 secretion system pilotin Guinier plot ln 8.60×100 Rg: 2.1 nm 0 (2.1 nm)-2 s2
(sRg)2I(s)/I(0)
Type 3 secretion system pilotin Kratky plot 1.104 0 3 sRg
p(r)
Type 3 secretion system pilotin pair distance distribution function Rg: 2.2 nm 0 Dmax: 8.0 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Type 3 secretion system pilotin DAMMIN model
Type 3 secretion system pilotin DAMFILT model

SAXS data from solutions of Type 3 secretion system pilotin, InvH70-147 in 20 mM HEPES, 150 mM NaCl, pH 7.5 were collected using a Rigaku BioSAXS-2000 instrument (University of British Columbia, Vancouver, Canada) at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2 and 16 mg/ml were measured at 6°C. Six successive 600 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Type 3 secretion system pilotin (InvH70)
Mol. type   Protein
Organism   Salmonella enterica subsp. enterica serovar Typhimurium
Olig. state   Dimer
Mon. MW   9.4 kDa
 
UniProt   P0CL43 (70-147)
Sequence   FASTA