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

SASDKF8 – N-Terminal Cytoplasmatic Region of the Translocated Intimin Receptor

Translocated intimin receptor Tir
MWexperimental 50 kDa
MWexpected 50 kDa
VPorod 46 nm3
log I(s) 6.22×10-2 6.22×10-3 6.22×10-4 6.22×10-5
Translocated intimin receptor Tir small angle scattering data  s, nm-1
ln I(s)
Translocated intimin receptor Tir Guinier plot ln 6.23×10-2 Rg: 3.5 nm 0 (3.5 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: 3.7 nm 0 Dmax: 14 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Synchrotron SAXS data from N-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 2.6942 m and at a wavelength of λ = 0.0947 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 20 mg/ml under a flow rate of 0.16 ml/min onto Shodex KW403-4F column (Showa Denko F.F.) at 10°C collecting 915 successive 2-second frames. The scattering intensities from N-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 0.15 < s <4.01 nm-1. The Rg values were estimated by applying the Guinier approximation in the range s < 1.3/Rg

Translocated intimin receptor Tir (N-Tir)
Mol. type   Protein
Organism   Escherichia coli O127:H6 (strain E2348/69 / EPEC)
Olig. state   Dimer
Mon. MW   25.1 kDa
 
UniProt   B7UM99 (1-233)
Sequence   FASTA