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Rational design of biosensors by computational analysis

Jens Reiners.

SASDXF3 – RTX cytotoxin pro-MbxA without acylation (pro-MbxA)

RTX cytotoxin pro-MbxA without acylation
MWexperimental 105 kDa
MWexpected 100 kDa
VPorod 154 nm3
log I(s) 4.90×10-2 4.90×10-3 4.90×10-4 4.90×10-5
RTX cytotoxin pro-MbxA without acylation small angle scattering data  s, nm-1
ln I(s)
RTX cytotoxin pro-MbxA without acylation Guinier plot ln 4.91×10-2 Rg: 3.8 nm 0 (3.8 nm)-2 s2
(sRg)2I(s)/I(0)
RTX cytotoxin pro-MbxA without acylation Kratky plot 1.104 0 √3 sRg
p(r)
RTX cytotoxin pro-MbxA without acylation pair distance distribution function Rg: 3.9 nm 0 Dmax: 13.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
RTX cytotoxin pro-MbxA without acylation ALPHAFOLD model
log I(s)
 s, nm-1
RTX cytotoxin pro-MbxA without acylation SREFLEX model
log I(s)
 s, nm-1
RTX cytotoxin pro-MbxA without acylation GASBOR model
Synchrotron SAXS data from solutions of RTX cytotoxin pro-MbxA without acylation (pro-MbxA) in 50 mM Tris, 100 mM NaCl, 10mM CaCl2, pH 7.8 were collected on the EMBL P12 beam line at the PETRA III storage ring (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.155 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 100.00 μl sample at 5.3 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superose 6 Increase 10/300 column at 20°C. 3000 successive 0.995 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.

RTX cytotoxin pro-MbxA without acylation (pro-MbxA)
Mol. type   Protein
Organism   Moraxella bovis
Olig. state   Monomer
Mon. MW   99.7 kDa
 
UniProt   Q93GI2
Sequence   FASTA