Novel lipid-interaction motifs within the C-terminal domain of Septin10 from Schistosoma mansoni

Cavini I, Fontes M, Zeraik A, Lopes J, Araújo A, Biochimica et Biophysica Acta (BBA) - Biomembranes :184371 (2024) DOI

SASDUV2 – MBP-fused Schistosoma mansoni Septin-10 C-terminal coiled-coil domain (MBP-SmSEPT10C)

Septin-10
MWexperimental 109 kDa
MWexpected 110 kDa
VPorod 175 nm3
log I(s) 3.22×10-2 3.22×10-3 3.22×10-4 3.22×10-5
Septin-10 small angle scattering data  s, nm-1
ln I(s)
Septin-10 Guinier plot ln 3.23×10-2 Rg: 5.7 nm 0 (5.7 nm)-2 s2
(sRg)2I(s)/I(0)
Septin-10 Kratky plot 1.104 0 √3 sRg
p(r)
Septin-10 pair distance distribution function Rg: 6.7 nm 0 Dmax: 27.4 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Septin-10 OMEGAFOLD model
log I(s)
 s, nm-1
Septin-10 ALPHAFOLD model
Septin-10 OMEGAFOLD model
log I(s)
 s, nm-1
Septin-10 DAMMIN model
Synchrotron SAXS data from solutions MBP-SmSEPT10C in 50 mM Tris, 150 mM NaCl, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.09464 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 45.00 μl sample at 5 mg/ml was injected at a 0.08 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 15°C. 600 successive 3 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 modelling presented are (from top to bottom): 1) MultiFoXS using parallel coiled-coil model; 2) MultiFoXS using parallel and antiparallel coiled-coil models (79:21 weight); 3) DAMMIN bead model (in P1 symmetry).

Septin-10 (MBP-SmSEPT10C)
Mol. type   Protein
Organism   Schistosoma mansoni
Olig. state   Dimer
Mon. MW   55.0 kDa
 
UniProt   G4VFI8 (307-412)
Sequence   FASTA