Small-angle X-ray and neutron scattering demonstrates that cell-free expression produces properly formed disc-shaped nanolipoprotein particles.

Cleveland TE 4th, He W, Evans AC, Fischer NO, Lau EY, Coleman MA, Butler P, Protein Sci 27(3):780-789 (2018) Europe PMC

SASDCF9 – Nanolipoprotein Particle (NLP) from Cell-Free Expression - 100% D2O SANS

Apolipoprotein A-I
MWexperimental 142 kDa
MWexpected 144 kDa
log I(s) 6.00×10-1 6.00×10-2 6.00×10-3 6.00×10-4
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I small angle scattering data  s, nm-1
ln I(s)
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I Guinier plot ln 6×10-1 Rg: 3.5 nm 0 (3.5 nm)-2 s2
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I Kratky plot 1.104 0 3 sRg
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I pair distance distribution function Rg: 3.4 nm 0 Dmax: 12.5 nm

Data validation

There are no models related to this curve.

Small-angle neutron scattering (SANS) data from solutions of Nanolipoprotein Particle (NLP) from Cell-Free Expression in PBS constituted in 100% D2O, pD 7.4, were collected on the NG7 instrument at the NIST Center for Neutron Research (Gaithersburg, MD, USA) using a 640x640 mm 3He position sensitive Ordela 2660N detector at a wavelength of λ = 0.6 nm (I(s) vs s, where s = 4πsinθ/λ and 2θ is the scattering angle). One solute concentration of 0.90 mg/ml was measured at 20°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Additional SANS measurement parameters. Three detector positions (in m) and count times (s) were employed: 1 m (two frames of 300 + 900 = 1200 s total count time); 4 m (two frames of 1800 + 1800 = 3600 s total count time); 13 m (two frames of 300 + 1200 = 1500 s total count time).

1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)
Mol. type   Other
Olig. state   Other
Mon. MW   0.7 kDa
Chemical formula
Apolipoprotein A-I (ApoA-I)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Dimer
Mon. MW   26.1 kDa
UniProt   Q00623 (67-264)
Sequence   FASTA