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

SASDHQ7 – Haloalkane dehalogenase variant DhaA115 - dimeric fraction

Haloalkane dehalogenase variant DhaA115 - dimeric fraction

MW^experimental	49	kDa
MW^expected	69	kDa
V^Porod	78	nm³

log I(s) 1.68×10⁰ 1.68×10^-1 1.68×10^-2 1.68×10^-3

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction small angle scattering data$ s, nm^-1

Log plot Log-Log plot

ln I(s)

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction Guinier plot$ ln 1.68×10⁰ R_g: 2.9 nm 0 (2.9 nm)^-2 s²

(sR_g)²I(s)/I(0)

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction Kratky plot$ 1.104 0 √3 sR_g

p(r)	$Haloalkane dehalogenase variant DhaA115 - dimeric fraction pair distance distribution function$ R_g: 3.1 nm 0 D_max: 8.9 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.6

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.948
1.386

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction CORAL model$

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction CORAL model$

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

$Haloalkane dehalogenase variant DhaA115 - dimeric fraction CORAL model$

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 8.7 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 - dimeric fraction (DhaA115 - dimeric)
Mol. type		Protein
Organism		Rhodococcus rhodochrous
Olig. state		Dimer
Mon. MW		34.4 kDa
Sequence		FASTA

PDB ID		6TY7

PDB ID		6XT8

PDB ID		6XTC