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

SASDCE9 – Nanolipoprotein Particle (NLP) with Telodendrimer - SAXS

1,2-dimyristoyl-sn-glycero-3-phosphocholine
Apolipoprotein A-I
MWexperimental 142 kDa
MWexpected 144 kDa
log I(s) 1.50×10-2 1.50×10-3 1.50×10-4 1.50×10-5
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 1.50×10-2 Rg: 4.4 nm 0 (4.4 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) with Telodendrimer 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 0.83 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