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

SASDCU7 – TRIP18(7.5R)SN-f5

TRIP18(7.5R)SN-f5

MW^I(0)	92	kDa
MW^expected	82	kDa
V^Porod	210	nm³

log I(s) 1.25×10⁴ 1.25×10³ 1.25×10² 1.25×10¹

TRIP18(7.5R)SN-f5 small angle scattering data

s, nm^-1

Log plot Log-Log plot

ln I(s)

ln 1.25×10⁴ R_g: 4.2 nm 0 (4.2 nm)^-2 s²

(sR_g)²I(s)/I(0)

1.104 0 √3 sR_g

p(r)	R_g: 4.2 nm 0 D_max: 14.5 nm

Data validation

Experimental data range

p(r) fit to data

Metric

Value

s_min D_max / π

0.7

N_Shannon

Worse

Better

Metric

Value

p_cormap

χ²

0.040
0.786

Worse

Better

Fits and models

log I(s)	s, nm^-1
+3 Δ/σ -3	s, nm^-1

Synchrotron SAXS data from solutions of TRIP18(7.5R)SN-f5 in 20 mM Tris 150 mM NaCl 10% glycerol, pH 7.5 were collected on the P12 beam line at the PETRA III storage ring (Hamburg, Germany) using a Pilatus 1M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). Solute concentrations ranging between 0.4 and 4.1 mg/ml were measured at 20°C. 20 successive 0.045 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 and the different curves were scaled for protein concentration. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve. To detect any potential concentration dependent effects a series consisting of 3 concentrations was measured. The highest concentration was 4.1 mg/ml, while the lowest concentrations was 0.4 mg/ml. Frames were carefully inspected for radiation damage and those not showing any radiation damage were then averaged. Before and after each sample scattering, matching buffer was measured and subtracted from sample scattering. Scattering curves were merged and analyzed using PRIMUS software.

TRIP18(7.5R)SN-f5 (TRIP18SN)
Mol. type		Protein
Organism		synthetic construct
Olig. state		Monomer
Mon. MW		81.9 kDa
Sequence		FASTA