Metabolic labeling: Taking advantage of bacterial pathways to prepare spectroscopically useful isotope patterns in proteins and nucleic acids
Year: 2008
Type: article
Abstract: Uniform labeling with the stable isotopes 15N, 13C, and/or 2H is a central enabling technology for NMR studies of proteins and nucleic acids. As the field advances into structural studies of larger systems and a more detailed characterization of molecular dynamics; however, there is an increasing need for schemes that specifically introduce labels at selected molecular sites only. For many purposes, knowledge of bacterial biosynthetic pathways allows procedures to be developed that produce desired labeling patterns directly in the expression culture, rather than via laborious chemical synthesis. In this work, we review the principles and applications of such “metabolic labeling” schemes. After an overview of key metabolic and biosynthetic processes in E. coli, we discuss pioneering applications of metabolic labeling and conclude with a detailed description of the two most prominent current uses of such techniques: The introduction of hydrogen or carbon isotopes specifically at side‐chain methyl groups of proteins and the removal of one‐bond 13C‐13C dipolar couplings for spin relaxation or solid‐state structural studies via alternate‐site labeling schemes. © 2008 Wiley Periodicals, Inc. Concepts Magn Reson Part A 32A: 34–55, 2008.
DOI: 10.1002/cmr.a.20103
Authors: Charles G. Hoogstraten, James E. Johnson
Institution: Michigan State University
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: bronze
OpenAlex ID: W2029656409