Inhibition of aggregation and biofilm formation by Uropathogenic Escherichia coli

Andrew Whitten.

SASDF33 – Antigen 43/Fragment antigen-binding region Fab10C12 complex (Ag43-Fab)

Alpha domain of Ag43a
Fragment antigen-binding region Fab10C12
MWI(0) 92 kDa
MWexpected 96 kDa
VPorod 103 nm3
log I(s) 2.17×10-2 2.17×10-3 2.17×10-4 2.17×10-5
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 small angle scattering data  s, nm-1
ln I(s)
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 Guinier plot ln 2.17×10-2 Rg: 4.3 nm 0 (4.3 nm)-2 s2
(sRg)2I(s)/I(0)
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 Kratky plot 1.104 0 √3 sRg
p(r)
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 pair distance distribution function Rg: 4.3 nm 0 Dmax: 15 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 DAMFILT model
log I(s)
 s, nm-1
Alpha domain of Ag43a Fragment antigen-binding region Fab10C12 CUSTOM IN-HOUSE model
X-ray synchrotron radiation scattering data from the from the Ag43-Fab complex in 25 mM HEPES 150mM NaCl pH 7.0 were collected on the SAXS/WAXS beam line of the Australian Synchrotron (Melbourne, Australia) in a SEC-SAXS with sheath flow configuration using a 2D Photon counting Pilatus 1M-W pixel detector (I(s) vs s, where s = 4π sin θ/λ and 2θ is the scattering angle; λ=0.10332 nm). Approximately 50 uL of a 5.0 mg/ml protein solution was injected onto a S200 Increase 5/150 column at a flow rate of 0.4 ml/min. Three hundred successive 1 second frames were collected during the SEC-SAXS run. The peak of the elution profile occured at ~3 mins after the start of the run, and the protein scattering was taken as the average of 14 frames collected 190-203 seconds after the peak eluted. The concentration range corresponding to the averaged frames was ~0.25 - 1.20 mg/ml, and the average concentration was taken as ~0.59 mg/ml. The buffer was taken as the sum of 50 frames collected between 1-2 mins after the start of the run. The data were normalized to the intensity of the transmitted beam and radially averaged and the scattering of the solvent-blank was subtracted. The models and corresponding fits include those derived from dummy-atom modelling using DAMMIN (top) and CRYSOL using the crystal structure (bottom).

SEC column = UNKNOWN. Sample injection volume = UNKNOWN. Flow rate = UNKNOWN

Alpha domain of Ag43a (Ag43)
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   49.1 kDa
Sequence   FASTA
 
Fragment antigen-binding region Fab10C12 (Fab10C12)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Monomer
Mon. MW   46.6 kDa
Sequence   FASTA