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Isotopic Labeling of Proteins in Halobacterium salinarum

Published

Author(s)

Thomas E. Cleveland, Zvi Kelman

Abstract

It is often necessary to obtain isotopically labeled proteins containing 15N, 13C or 2H for NMR; and 2H for small-angle neutron scattering (SANS) or neutron diffraction studies. To achieve uniform isotopic labeling, protein expression is most commonly performed in Escherichia coli or yeast using labeled media. However, proteins from extreme halophiles sometimes require a cellular environment with high ionic strength, and cannot be heterologously expressed in E. coli or yeast in functional form. We present here methods for the cultivation of Halobacterium salinarum in isotopically labeled rich media, using commercially available isotopically-labeled hydrolysates. The methods described here are both technically simple and relatively inexpensive.
Citation
Methods in Enzymology
Volume
565
Publisher Info
Elsevier, Oxford, -1

Keywords

archaea, protein labeling

Citation

Cleveland, T. and Kelman, Z. (2015), Isotopic Labeling of Proteins in Halobacterium salinarum, Methods in Enzymology, Elsevier, Oxford, -1, [online], https://doi.org/10.1016/bs.mie.2015.06.002 (Accessed April 22, 2024)
Created August 4, 2015, Updated November 10, 2018