Identity Determinants of the Translocation Signal for a Type 1 Secretion System

Spitz O, Erenburg I, Kanonenberg K, Peherstorfer S, Lenders M, Reiners J, Ma M, Luisi B, Smits S, Schmitt L, Frontiers in Physiology 12 (2022) DOI

SASDM67 – Hemolysin A (HlyA) from Escherichia coli UTI89

Hemolysin, plasmid (Hemolysin A)
MWexperimental 208 kDa
MWexpected 220 kDa
VPorod 346 nm3
log I(s) 9.59×101 9.59×100 9.59×10-1 9.59×10-2
Hemolysin, plasmid (Hemolysin A) small angle scattering data  s, nm-1
ln I(s)
Hemolysin, plasmid (Hemolysin A) Guinier plot ln 9.59×101 Rg: 6.7 nm 0 (6.7 nm)-2 s2
(sRg)2I(s)/I(0)
Hemolysin, plasmid (Hemolysin A) Kratky plot 1.104 0 √3 sRg
p(r)
Hemolysin, plasmid (Hemolysin A) pair distance distribution function Rg: 7.0 nm 0 Dmax: 25.3 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Hemolysin, plasmid (Hemolysin A) GASBOR model
log I(s)
 s, nm-1
Hemolysin, plasmid (Hemolysin A) ALPHAFOLD model
Synchrotron SAXS data from solutions of Hemolysin A in 100 mM HEPES pH 8.0, 250 mM NaCl, 10 mM CaCl2 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 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 110.00 μl sample at 8 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superose 6 Increase 10/300 column at 10°C. 1500 successive 2 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 from those sample frames encompassing the SEC elution peak.

Hemolysin, plasmid (Hemolysin A) (HlyA)
Mol. type   Protein
Organism   Escherichia coli UTI89
Olig. state   Dimer
Mon. MW   110.2 kDa
 
UniProt   P08715 (1-1024)
Sequence   FASTA