The pathogen-encoded signalling receptor Tir exploits host-like intrinsic disorder for infection.

Vieira MFM, Hernandez G, Zhong Q, Arbesú M, Veloso T, Gomes T, Martins ML, Monteiro H, Frazão C, Frankel G, Zanzoni A, Cordeiro TN, Commun Biol 7(1):179 (2024) Europe PMC

SASDKG8 – Central domain of the N-terminal Region of the Translocated Intimin Receptor (60-200)

Translocated intimin receptor Tir
MWexperimental 31 kDa
MWexpected 32 kDa
VPorod 47 nm3
log I(s) 4.28×10-2 4.28×10-3 4.28×10-4 4.28×10-5
Translocated intimin receptor Tir small angle scattering data  s, nm-1
ln I(s)
Translocated intimin receptor Tir Guinier plot ln 4.28×10-2 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
Translocated intimin receptor Tir Kratky plot 1.104 0 √3 sRg
p(r)
Translocated intimin receptor Tir pair distance distribution function Rg: 3.6 nm 0 Dmax: 13 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Translocated intimin receptor Tir GASBOR model
Synchrotron SAXS data from NS-Tir in 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM DTT, pH 8.0 were collected on the B21 beam-line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 2.696 m and at a wavelength of λ = 0.0946 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed using an Agilent 1200 HPLC System by injecting a 60 μl sample at 25 mg/ml under a flow rate of 0.16 ml/min onto Shodex KW402.5-4F column (Showa Denko F.F.) at 10°C collecting 915 successive 2-second frames. The scattering intensities from NS-Tir elution peak region (35-frames) were integrated, buffer subtracted and averaged using the ScÅtter software to produce the averaged SEC-SAXS profile. From this profile, the pair-wise distance distribution function, P(r), was obtained by indirect Fourier Transform with GNOM using a momentum transfer range of 0.09 < s <3.59 nm-1. The Rg values were estimated by applying the Guinier approximation in the range s < 1.3/Rg. The displayed SAXS-generated ab initio molecular reconstruction was obtained using GASBOR.

Translocated intimin receptor Tir (NS-Tir)
Mol. type   Protein
Organism   Escherichia coli O127:H6 (strain E2348/69 / EPEC)
Olig. state   Dimer
Mon. MW   15.8 kDa
 
UniProt   B7UM99 (60-200)
Sequence   FASTA