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

SASDCD9 – Nanolipoprotein Particle (NLP) from Cell-Free Expression - SAXS

1,2-dimyristoyl-sn-glycero-3-phosphocholine
Apolipoprotein A-I
MWexperimental 142 kDa
MWexpected 144 kDa
log I(s) 1.90×10-2 1.90×10-3 1.90×10-4 1.90×10-5
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 1.90×10-2 Rg: 4.7 nm 0 (4.7 nm)-2 s2
(sRg)2I(s)/I(0)
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I Kratky plot 1.104 0 √3 sRg
p(r)
1,2-dimyristoyl-sn-glycero-3-phosphocholine Apolipoprotein A-I pair distance distribution function Rg: 4.7 nm 0 Dmax: 12.7 nm

Data validation

There are no models related to this curve.

SAXS data from solutions of Nanolipoprotein Particle (NLP) from Cell-Free Expression in PBS constituted in D2O, pD 7.4, were collected on the NIST/IBBR SAXS instrument (Rockville, United States) using a PILATUS 300K detector at a wavelength of λ = 0.15418 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 SAXS measurement parameters. Three detector positions (in m) and exposure times (s) were employed: 0.355 m (18 frames of 600 s = 10800 s total exposure time); 0.705 m (18 frames of 600 s = 10800 s total exposure time); 1.305 m (18 frames of 600 s = 10800 s total exposure 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