title: Isotopic labeling of proteins in Halobacterium salinarum.
abstract: It is often necessary to obtain isotopically labeled proteins containing (15)N, (13)C, or (2)H for nuclear magnetic resonance; and (2)H for small-angle neutron scattering 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.
authors.affiliation: NIST Center for Neutron Research, National Institute of Standards and Technology, Gaithersburg, Maryland, USA; Biomolecular Structure and Function Group, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, Maryland, USA. Electronic address: thomas.cleveland@nist.gov.
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Cleveland
authors.affiliation: Biomolecular Labeling Laboratory, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, Maryland, USA; Biomolecular Structure and Function Group, Institute for Bioscience and Biotechnology Research, National Institute of Standards and Technology and the University of Maryland, Rockville, Maryland, USA.
authors.forename: Zvi
authors.initials: Z
authors.lastname: Kelman
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry*; D019617:Halobacterium salinarum / Q000737:chemistry* / Q000254:growth & development; D007553:Isotope Labeling; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D019843:Archaeal Proteins
keywords: Algal hydrolysate; Archaea; Bacteriorhodopsin; Deuteration; Isotope labeling; Protein expression; Protein labeling