Structural models of intrinsically disordered and calcium-bound folded states of a protein adapted for secretion.

O'Brien DP, Hernandez B, Durand D, Hourdel V, Sotomayor-Pérez AC, Vachette P, Ghomi M, Chamot-Rooke J, Ladant D, Brier S, Chenal A, Sci Rep 5:14223 (2015) Europe PMC

SASDGM5 – Holo-RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA)

RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA)
MWI(0) 76 kDa
MWexpected 73 kDa
VPorod 89 nm3
log I(s) 2.53×10-2 2.53×10-3 2.53×10-4 2.53×10-5
RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) small angle scattering data  s, nm-1
ln I(s)
RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) Guinier plot ln 2.53×10-2 Rg: 4.4 nm 0 (4.4 nm)-2 s2
(sRg)2I(s)/I(0)
RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) Kratky plot 1.104 0 3 sRg
p(r)
RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) pair distance distribution function Rg: 4.5 nm 0 Dmax: 15.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) OTHER model

Synchrotron SAXS data from solutions of the holo-RD domain of B. Pertussis adenylate cyclase toxin (CyaA) in 20 mM Hepes, 150 mM NaCl, 2 mM DTT, 4 mM CaCl2, pH 7.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.1033 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 8 mg/ml was injected at a 0.20 ml/min flow rate onto a Agilent Bio SEC-3, 300 Å column at 15°C. 250 successive 1 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 scattered intensities were displayed on an absolute scale (cm-1) using the scattering of water. Frames were examined individually and 10 identical frames obtained at the end of the elution peak were averaged and further processed. The corresponding concentration was 0.38g/L.

RD domain of B. Pertussis Adenylate Cyclase Toxin (CyaA) (RD domain of CyaA)
Mol. type   Protein
Organism   Bordetella pertussis
Olig. state   Monomer
Mon. MW   72.6 kDa
 
UniProt   P0DKX7 (1007-1706)
Sequence   FASTA