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Small-Angle X-ray and Neutron Scattering Demonstrates that Cell-Free Expression Produces Properly Formed Disc-Shaped Nanolipoprotein Particles

Published

Author(s)

Thomas E Cleveland, Wei He, Angela C. Evans, Nicholas O. Fischer, Matthew A. Coleman, Paul Butler

Abstract

Nanolipoprotein particles (NLPs), composed of a membrane scaffold protein and lipids, have been used to support membrane proteins in a native-like bilayer environment for biochemical or 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 devloped 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 va detergent removal. 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.
Citation
Protein Science
Volume
27

Keywords

NLPs, nanodiscs, nanolipoprotein particles, SANS, small-angle neutron scattering, cell-free expression

Citation

, T. , He, W. , , A. , , N. , , M. and Butler, P. (2017), Small-Angle X-ray and Neutron Scattering Demonstrates that Cell-Free Expression Produces Properly Formed Disc-Shaped Nanolipoprotein Particles, Protein Science, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=922386 (Accessed February 28, 2024)
Created December 20, 2017, Updated June 26, 2018