Activity of lymphostatin, a lymphocyte inhibitory virulence factor of pathogenic Escherichia coli, is dependent on a cysteine protease motif.

Bease AG, Blackburn EA, Chintoan-Uta C, Webb S, Cassady-Cain RL, Stevens MP, J Mol Biol :167200 (2021) Europe PMC

SASDKP9 – Lymphostatin C1480A mutant pH5.5

Efa1/LifA protein (C1480A mutant)
MWexperimental 403 kDa
MWexpected 367 kDa
VPorod 560 nm3
log I(s) 1.36×10-1 1.36×10-2 1.36×10-3 1.36×10-4
Efa1/LifA protein (C1480A mutant) small angle scattering data  s, nm-1
ln I(s)
Efa1/LifA protein (C1480A mutant) Guinier plot ln 1.36×10-1 Rg: 5.8 nm 0 (5.8 nm)-2 s2
(sRg)2I(s)/I(0)
Efa1/LifA protein (C1480A mutant) Kratky plot 1.104 0 √3 sRg
p(r)
Efa1/LifA protein (C1480A mutant) pair distance distribution function Rg: 5.9 nm 0 Dmax: 19.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Efa1/LifA protein (C1480A mutant) DAMFILT model
log I(s)
 s, nm-1
Synchrotron SAXS data of C1480A single point mutant Lymphostatin in 50 mM Bis-Tris, pH 5.5; 100 mM NaCl; 3% glycerol (v/v) were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector 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 using a 2.4 mL superdex200 3.2/300 chromatography column (GE healthcare). 35 μl of mutant lymphostatin solution at 1.93 mg/ml was injected at 0.075 ml/min and 620 successive 1.5 second data-frames were collected at 15°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the eluted buffer with no macromolecular component was subtracted.

Efa1/LifA protein (C1480A mutant) (C1480A Lymphostatin)
Mol. type   Protein
Organism   Escherichia coli O127:H6 (strain E2348/69 / EPEC)
Olig. state   Monomer
Mon. MW   366.7 kDa
 
UniProt   B7UI23 (1-3223)
Sequence   FASTA