Design of coiled-coil protein-origami cages that self-assemble in vitro and in vivo.

Ljubetič A, Lapenta F, Gradišar H, Drobnak I, Aupič J, Strmšek Ž, Lainšček D, Hafner-Bratkovič I, Majerle A, Krivec N, Benčina M, Pisanski T, Veličković TĆ, Round A, Carazo JM, Melero R, Jerala R, Nat Biotechnol 35(11):1094-1101 (2017) Europe PMC

SASDCX7 – TET12(1.10)S-f5b

TET12(1.10)S-f5b
MWI(0) 47 kDa
MWexpected 53 kDa
VPorod 127 nm3
log I(s) 3.12×102 3.12×101 3.12×100 3.12×10-1
TET12(1.10)S-f5b small angle scattering data  s, nm-1
ln I(s)
TET12(1.10)S-f5b Guinier plot ln 3.13×102 Rg: 3.4 nm 0 (3.4 nm)-2 s2
(sRg)2I(s)/I(0)
TET12(1.10)S-f5b Kratky plot 1.104 0 √3 sRg
p(r)
TET12(1.10)S-f5b pair distance distribution function Rg: 3.4 nm 0 Dmax: 11.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
TET12(1.10)S-f5b MODELLER model
Synchrotron SAXS data from solutions of TET12(1.10)S-f5b in 20 mM Tris, 150 mM NaCl, 10% glycerol, pH 7.5, were collected at the BM29 beam line at the ESRF storage ring (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.89 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, where 2θ is the scattering angle). A solute concentration series ranging between 1 and 7 mg/ml were measured at 20°C. For each sample, data was collected over 10 frames lasting 2 s. The data were normalized to the intensity of the transmitted beam and radially averaged. Each individual frames were carefully inspected for radiation damage and those not showing any radiation damage were then averaged. The scattering of the matched solvent-blank was subtracted and the different curves were scaled for protein concentration. The protein did not exhibit any concentration dependent effects, therefore the curve measured for the sample with the highest concentration was used for subsequent analysis. Scattering curves were analyzed using PRIMUS software.

TET12(1.10)S-f5b
Mol. type   Protein
Organism   synthetic construct
Olig. state   Monomer
Mon. MW   53.2 kDa
Sequence   FASTA