Continuous Assembly of β-Roll Structures Is Implicated in the Type I-Dependent Secretion of Large Repeat-in-Toxins (RTX) Proteins.

Motlova L, Klimova N, Fiser R, Sebo P, Bumba L, J Mol Biol (2020) Europe PMC

SASDJU3 – Adenylate cyclase toxin CyaA RTX block IV-V

Bifunctional hemolysin/adenylate cyclase
MWI(0) 35 kDa
MWexpected 32 kDa
VPorod 45 nm3
log I(s) 4.60×103 4.60×102 4.60×101 4.60×100
Bifunctional hemolysin/adenylate cyclase small angle scattering data  s, nm-1
ln I(s)
Bifunctional hemolysin/adenylate cyclase Guinier plot ln 4.61×103 Rg: 2.7 nm 0 (2.7 nm)-2 s2
Bifunctional hemolysin/adenylate cyclase Kratky plot 1.104 0 3 sRg
Bifunctional hemolysin/adenylate cyclase pair distance distribution function Rg: 2.6 nm 0 Dmax: 8.6 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Bifunctional hemolysin/adenylate cyclase DAMMIF model

Synchrotron SAXS data from solutions of CyaA RTX block IV-V in 10 mM Tris HCl, 150 mM NaCl, 10 mM CaCl₂, pH 8 were collected on the EMBL P12 beam line at PETRA III (DESY, Hamburg, Germany) using a Pilatus 2M detector at a sample-detector distance of 3.1 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.8 and 6.8 mg/ml were measured at 10°C. 20 successive 0.050 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 concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.

The ab initio model displayed is the likely bead- and volume-occupancy corrected representation based on the spatial alignment of an individual model cohort (damfilt model) and may not necessarily reflect the displayed model fit to the data.

Bifunctional hemolysin/adenylate cyclase (CyaA)
Mol. type   Protein
Organism   Bordetella pertussis
Olig. state   Monomer
Mon. MW   31.8 kDa
UniProt   P0DKX7 (1372-1681)
Sequence   FASTA