Preparation of a Deuterated Membrane Protein for Small-Angle Neutron Scattering

Book · Thu Apr 01 00:00:00 EDT 2021

OSTI ID:1807219

Wu, Yuqi ^[1]; Weiss, Kevin L. ^[2]; Lieberman, Raquel ^[1]

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1. Georgia Tech
2. ORNL

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.

• Research Organization: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
• Sponsoring Organization: USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
• DOE Contract Number: AC05-00OR22725
• OSTI ID: 1807219
• Country of Publication: United States
• Language: English