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TitleSmall-angle X-ray and neutron scattering demonstrates that cell-free expression produces properly formed disc-shaped nanolipoprotein particles.
Journal, issue, pagesProtein Sci, Vol. 27, Issue 3, Page 780-789, Year 2018
Publish dateFeb 13, 2018
AuthorsThomas E Cleveland / Wei He / Angela C Evans / Nicholas O Fischer / Edmond Y Lau / Matthew A Coleman / Paul Butler /
PubMed AbstractNanolipoprotein particles (NLPs), composed of membrane scaffold proteins and lipids, have been used to support membrane proteins in a native-like bilayer environment for biochemical and structural ...Nanolipoprotein particles (NLPs), composed of membrane scaffold proteins and lipids, have been used to support membrane proteins in a native-like bilayer environment for biochemical and structural studies. Traditionally, these NLPs have been prepared by the controlled removal of detergent from a detergent-solubilized protein-lipid mixture. Recently, an alternative method has been developed using direct cell-free expression of the membrane scaffold protein in the presence of preformed lipid vesicles, which spontaneously produces NLPs without the need for detergent at any stage. Using SANS/SAXS, we show here that NLPs produced by this cell-free expression method are structurally indistinguishable from those produced using detergent removal methodologies. This further supports the utility of single step cell-free methods for the production of lipid binding proteins. In addition, detailed structural information describing these NLPs can be obtained by fitting a capped core-shell cylinder type model to all SANS/SAXS data simultaneously.
External linksProtein Sci / PubMed:29266475 / PubMed Central
MethodsSAS (neutron source)
Structure data

SASDC99: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS
Method: SAXS/SANS

SASDCB9: Nanolipoprotein Particle (NLP) assembled with tail-deuterated DMPC - 100% D2O SANS
Method: SAXS/SANS

SASDCC9: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS
Method: SAXS/SANS

SASDCF9: Nanolipoprotein Particle (NLP) from Cell-Free Expression - 100% D2O SANS
Method: SAXS/SANS

SASDCG9: Nanolipoprotein Particle (NLP) with Telodendrimer - 100% D2O SANS
Method: SAXS/SANS

Source
  • Mus musculus (house mouse)

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