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

Authors

Kiyoshi Ozawa, University of WollongongFollow
Nicholas E. Dixon, University of WollongongFollow
Gottfried Otting, Australian National University

RIS ID

16946

Publication Details

Ozawa, K., Dixon, N. E. & Otting, G. (2005). Cell-free synthesis of 15N-labeled proteins for NMR studies. IUBMB Life, 57 (9), 615-622.

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 N-15-labeled proteins from N-15-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 N-15-labeled proteins for analysis by NMR spectroscopy.