Cell-free synthesis of 15N-labeled proteins for NMR studies

Kiyoshi Ozawa, Nicholas E. Dixon, Gottfried Otting*

*Corresponding author for this work

    Research output: Contribution to journalReview articlepeer-review

    30 Citations (Scopus)

    Abstract

    Modern cell-free in vitro protein synthesis systems present powerful tools for the synthesis of isotope-labeled proteins in high yields. The production of selectively 15N-labeled proteins from 15N-labeled amino acids is particularly economic and yields are often sufficient to analyze the proteins very quickly by two-dimensional NMR spectra recorded of the crude reaction mixture without concentration or chromatographic purification of the protein. We review methodological aspects of cell-free in vitro protein synthesis based on an Escherichia coli cell extract, in particular with regard to the production of 15N-labeled proteins for analysis by NMR spectroscopy.

    Original languageEnglish
    Pages (from-to)615-622
    Number of pages8
    JournalIUBMB Life
    Volume57
    Issue number9
    DOIs
    Publication statusPublished - Sept 2005

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