Preparation of a Deuterated Membrane Protein for Small-Angle Neutron Scattering
Yuqi Wu1, Kevin L Weiss2, Raquel L Lieberman3
1School of Chemistry & Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA.
Abstract:
This chapter outlines a protocol developed to prepare a purified deuterated membrane protein for a small-angle neutron scattering (SANS) experiment. SANS is a noninvasive technique well suited to studying membrane protein solution structures, and deuteration enhances the signal from the protein over the background (Breyton et al., Eur Phys J E Soft Matter 36 (7):71, 2013; Garg et al., Biophys J 101 (2):370-377, 2011). We present our workflow: transformation of our plasmid into E. coli, cell growth and expression of our deuterated protein, membrane isolation, detergent solubilization, protein purification, purity assessment, and final preparation for SANS.
