Towards improved understanding of intersubunit interactions in modular polyketide biosynthesis: docking in the enacyloxin IIa polyketide synthase.

Risser F, Collin S, Dos Santos-Morais R, Gruez A, Chagot B, Weissman KJ, J Struct Biol :107581 (2020) Europe PMC

SASDJH2 – Beta-ketoacyl synthase (Bamb_5924) N-terminal docking domain mutant K32A

Beta-ketoacyl synthase Bamb_5924, K32A mutant
MWexperimental 10 kDa
MWexpected 10 kDa
VPorod 22 nm3
log I(s) 2.68×10-2 2.68×10-3 2.68×10-4 2.68×10-5
Beta-ketoacyl synthase Bamb_5924, K32A mutant small angle scattering data  s, nm-1
ln I(s)
Beta-ketoacyl synthase Bamb_5924, K32A mutant Guinier plot ln 2.68×10-2 Rg: 1.8 nm 0 (1.8 nm)-2 s2
Beta-ketoacyl synthase Bamb_5924, K32A mutant Kratky plot 1.104 0 3 sRg
Beta-ketoacyl synthase Bamb_5924, K32A mutant pair distance distribution function Rg: 1.9 nm 0 Dmax: 8 nm

Data validation

There are no models related to this curve.

Synchrotron SAXS data from solutions of Bamb_5924 NDD mutant K32A in 20 mM Tris-HCl, 200 mM NaCl, 5% glycerol, pH 7.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a sample-detector distance of 1.8 m and 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. The SEC parameters were as follows: A 50.00 μl sample at 28 mg/ml was injected at a 0.10 ml/min flow rate onto a Agilent Bio SEC-3, 100 Å column at 15°C. 100 successive 0.750 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 protein samples were injected using the online automatic sample changer into a pre-equilibrated HPLC-coupled size-exclusion chromatography column.

Beta-ketoacyl synthase Bamb_5924, K32A mutant (Bamb_5924 NDD K32A)
Mol. type   Protein
Organism   Burkholderia ambifaria
Olig. state   Dimer
Mon. MW   4.9 kDa
UniProt   Q0B304 (1-41)
Sequence   FASTA