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

SASDCB9 – Nanolipoprotein Particle (NLP) assembled with tail-deuterated DMPC - 100% D2O SANS

Apolipoprotein A-I
1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine
MWexperimental 148 kDa
MWexpected 149 kDa
log I(s) 4.80×10-2 4.80×10-3 4.80×10-4 4.80×10-5
Apolipoprotein A-I 1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine small angle scattering data  s, nm-1
ln I(s)
Apolipoprotein A-I 1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine Guinier plot ln 4.80×10-2 Rg: 4.3 nm 0 (4.3 nm)-2 s2
(sRg)2I(s)/I(0)
Apolipoprotein A-I 1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine Kratky plot 1.104 0 3 sRg
p(r)
Apolipoprotein A-I 1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine pair distance distribution function Rg: 4.2 nm 0 Dmax: 10.7 nm

Data validation


There are no models related to this curve.

Small-angle neutron scattering (SANS) data from solutions of Nanolipoprotein Particle (NLP) assembled with tail-deuterated DMPC 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 0.88 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 600 + 600 = 1200 s total count time); 4 m (two frames of 1800 + 1800 = 3600 s total count time); 13 m (two frames of 600 + 600 = 1200 s total count time).

Apolipoprotein A-I (ApoA-I)
Mol. type   Protein
Organism   Mus musculus
Olig. state   Dimer
Mon. MW   25.2 kDa
 
UniProt   Q00623 (67-264)
Sequence   FASTA
 
1,2-dimyristoyl-d54-sn-glycero-3-phosphocholine (d54-DMPC)
Mol. type   Other
Olig. state   Other
Mon. MW   0.7 kDa
Chemical formula