+Search query
-Structure paper
Title | Small-angle X-ray and neutron scattering demonstrates that cell-free expression produces properly formed disc-shaped nanolipoprotein particles. |
---|---|
Journal, issue, pages | Protein Sci, Vol. 27, Issue 3, Page 780-789, Year 2018 |
Publish date | Feb 13, 2018 |
![]() | Thomas E Cleveland / Wei He / Angela C Evans / Nicholas O Fischer / Edmond Y Lau / Matthew A Coleman / Paul Butler / ![]() |
PubMed Abstract | 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 ...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. |
![]() | ![]() ![]() ![]() |
Methods | SAS (neutron source) |
Structure data | ![]() SASDC99: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 100% D2O SANS ![]() SASDCB9: Nanolipoprotein Particle (NLP) assembled with tail-deuterated DMPC - 100% D2O SANS ![]() SASDCC9: Nanolipoprotein Particle (NLP) from In Vitro Assembly - 42% D2O SANS ![]() SASDCF9: Nanolipoprotein Particle (NLP) from Cell-Free Expression - 100% D2O SANS ![]() SASDCG9: Nanolipoprotein Particle (NLP) with Telodendrimer - 100% D2O SANS |
Source |
|