Structure and dynamics of Toll immunoreceptor activation in the mosquito Aedes aegypti.

Saucereau Y, Wilson TH, Tang MCK, Moncrieffe MC, Hardwick SW, Chirgadze DY, Soares SG, Marcaida MJ, Gay NJ, Gangloff M, Nat Commun 13(1):5110 (2022) Europe PMC

SASDKY8 – Aedes aegypti Toll5A ectodomain in complex with Spz1C Cys-knot domain

Toll-like receptor
AAEL013433-PA (Cystine-knot_cytokine)
MWexperimental 270 kDa
MWexpected 201 kDa
VPorod 567 nm3
log I(s) 2.94×10-1 2.94×10-2 2.94×10-3 2.94×10-4
Toll-like receptor AAEL013433-PA (Cystine-knot_cytokine) small angle scattering data  s, nm-1
ln I(s)
Toll-like receptor AAEL013433-PA (Cystine-knot_cytokine) Guinier plot ln 2.94×10-1 Rg: 5.5 nm 0 (5.5 nm)-2 s2
(sRg)2I(s)/I(0)
Toll-like receptor AAEL013433-PA (Cystine-knot_cytokine) Kratky plot 1.104 0 3 sRg
p(r)
Toll-like receptor AAEL013433-PA (Cystine-knot_cytokine) pair distance distribution function Rg: 5.7 nm 0 Dmax: 21.5 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of the Aedes aegypti Toll5A ectodomain in complex with Spz1C Cys-knot domain in 50 mM NaCl, 50 mM Tris-HCl, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.1 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 55.00 μl sample at 5 mg/ml was injected at a 0.075 ml/min flow rate onto a GE Superdex 6 Increase 3.2/300 column at 20°C. 31 successive 3 second frames were collected through the SEC elution peak of the sample. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

The Toll5A-Spz1C complex was concentrated to 5 mg/mL in 50mM NaCl, 50 mM Tris-HCl pH7.5 and injected into a Superose 6 Increase 3.2 equilibrated in the same buffer at a flow rate of 0.075 ml/min. Data points were collected every 3 sec.

Toll-like receptor (T5A)
Mol. type   Protein
Organism   Aedes aegypti
Olig. state   Dimer
Mon. MW   87.6 kDa
 
UniProt   A0A6I8TEX2 (28-789)
Sequence   FASTA
 
AAEL013433-PA (Cystine-knot_cytokine)
Mol. type   Protein
Organism   Aedes aegypti
Olig. state   Dimer
Mon. MW   13.0 kDa
 
UniProt   Q16J57 (142-243)
Sequence   FASTA