SidF, a dual substrate N5-acetyl-N5-hydroxy-L-ornithine transacetylase involved in Aspergillus fumigatus siderophore biosynthesis

Poonsiri T, Stransky J, Demitri N, Haas H, Cianci M, Benini S, Journal of Structural Biology: X 11:100119 (2025) DOI

SASDV98 – Full-length N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF

N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF
MWexperimental 208 kDa
MWexpected 223 kDa
VPorod 412 nm3
log I(s) 8.54×10-1 8.54×10-2 8.54×10-3 8.54×10-4
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF small angle scattering data  s, nm-1
ln I(s)
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF Guinier plot ln 8.55×10-1 Rg: 4.0 nm 0 (4.0 nm)-2 s2
(sRg)2I(s)/I(0)
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF Kratky plot 1.104 0 √3 sRg
p(r)
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF pair distance distribution function Rg: 4.3 nm 0 Dmax: 19 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Full-length N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF Rg histogram Rg, nm
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF EOM/RANCH model
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF EOM/RANCH model
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF EOM/RANCH model
N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF EOM/RANCH model
SAXS data from solutions of full-length sidF in 50 mM Tris, 200 mM NaCl, pH 8 were collected using an Anton Paar SAXSpoint 2.0 instrument at the Institute of Biotechnology, Czech Academy of Sciences/Centre of Molecular Structure (Vestec, Czech Republic) equipped with an Eiger R 1M detector at a sample-detector distance of 0.8 m and at a wavelength of λ = 0.134 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 400.00 μl sample at 12.7 mg/ml was injected at a 0.05 ml/min flow rate onto a Cytiva Superdex 200 Increase 10/300 column at 20°C. 65 successive 15 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.

N(5)-hydroxyornithine:cis-anhydromevalonyl coenzyme A-N(5)-transacylase sidF (SidF)
Mol. type   Protein
Organism   Aspergillus fumigatus (strain ATCC MYA-4609 / CBS 101355 / FGSC A1100 / Af293)
Olig. state   Tetramer
Mon. MW   55.7 kDa
 
UniProt   Q4WF55 (1-462)
Sequence   FASTA