Cryo-EM structures of Trypanosoma brucei gambiense ISG65 with human complement C3 and C3b and their roles in alternative pathway restriction.

Sülzen H, Began J, Dhillon A, Kereïche S, Pompach P, Votrubova J, Zahedifard F, Šubrtova A, Šafner M, Hubalek M, Thompson M, Zoltner M, Zoll S, Nat Commun 14(1):2403 (2023) Europe PMC

SASDP99 – invariant surface glycoprotein ISG65

65 kDa invariant surface glycoprotein, putative
MWexperimental 40 kDa
MWexpected 41 kDa
VPorod 53 nm3
log I(s) 3.18×101 3.18×100 3.18×10-1 3.18×10-2
65 kDa invariant surface glycoprotein, putative small angle scattering data  s, nm-1
ln I(s)
65 kDa invariant surface glycoprotein, putative Guinier plot ln 3.18×101 Rg: 3.3 nm 0 (3.3 nm)-2 s2
(sRg)2I(s)/I(0)
65 kDa invariant surface glycoprotein, putative Kratky plot 1.104 0 3 sRg
p(r)
65 kDa invariant surface glycoprotein, putative pair distance distribution function Rg: 3.6 nm 0 Dmax: 12.5 nm

Data validation


Fits and models


log I(s)
 s, nm-1
invariant surface glycoprotein ISG65 Rg histogram Rg, nm
65 kDa invariant surface glycoprotein, putative EOM/RANCH model
65 kDa invariant surface glycoprotein, putative EOM/RANCH model
65 kDa invariant surface glycoprotein, putative EOM/RANCH model
65 kDa invariant surface glycoprotein, putative EOM/RANCH model
65 kDa invariant surface glycoprotein, putative EOM/RANCH model
65 kDa invariant surface glycoprotein, putative EOM/RANCH model

Synchrotron SAXS data from solutions of invariant surface glycoprotein ISG65 in 20 mM HEPES, 150 mM NaCl, 3% (v/v) glycerol, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus3 2M detector at a sample-detector distance of 2.8 m and at a wavelength of λ = 0.099 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 6.2 mg/ml was injected at a 0.08 ml/min flow rate onto a GE Superdex 200 Increase 3.2/300 column at 20°C. 1200 successive 1 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.

65 kDa invariant surface glycoprotein, putative
Mol. type   Protein
Organism   Trypanosoma brucei gambiense (strain MHOM/CI/86/DAL972)
Olig. state   Monomer
Mon. MW   40.8 kDa
 
UniProt   C9ZJ67 (18-363)
Sequence   FASTA