Decoding the intricate network of molecular interactions of a hyperstable engineered biocatalyst

Markova K, Chmelova K, Marques S, Carpentier P, Bednar D, Damborsky J, Marek M, Chemical Science 11(41):11162-11178 (2020) DOI

SASDHP7 – Haloalkane dehalogenase variant DhaA115 - monomeric fraction

Haloalkane dehalogenase variant DhaA115 -monomeric fraction
MWexperimental 22 kDa
MWexpected 34 kDa
VPorod 41 nm3
log I(s) 2.20×10-1 2.20×10-2 2.20×10-3 2.20×10-4
Haloalkane dehalogenase variant DhaA115 -monomeric fraction small angle scattering data  s, nm-1
ln I(s)
Haloalkane dehalogenase variant DhaA115 -monomeric fraction Guinier plot ln 2.20×10-1 Rg: 1.9 nm 0 (1.9 nm)-2 s2
(sRg)2I(s)/I(0)
Haloalkane dehalogenase variant DhaA115 -monomeric fraction Kratky plot 1.104 0 √3 sRg
p(r)
Haloalkane dehalogenase variant DhaA115 -monomeric fraction pair distance distribution function Rg: 1.9 nm 0 Dmax: 6.0 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Haloalkane dehalogenase variant DhaA115 -monomeric fraction PDB (PROTEIN DATA BANK) model
SAXS data from solutions of haloalkane dehalogenase variant DhaA115 in 50 mM potassium phosphate buffer (41 mM K₂HPO₄, 9mM KH₂PO₄), pH 7.5 were collected using a Rigaku BioSAXS-1000 instrument at CEITEC (Brno, Czech Republic) equipped with a Pilatus 100K detector at a sample-detector distance of 0.5 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 2.2 and 8.5 mg/ml were measured at 20°C. Six successive 300 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Haloalkane dehalogenase variant DhaA115 -monomeric fraction (DhaA115 - monomeric)
Mol. type   Protein
Organism   Rhodococcus rhodochrous
Olig. state   Monomer
Mon. MW   34.4 kDa
Sequence   FASTA
 
PDB ID   6SP5