Cell-free protein synthesis for analysis by NMR spectroscopy
Date
2008
Authors
Apponyi, Margit
Ozawa, Kiyoshi
Dixon, Nicholas
Otting, Gottfried
Journal Title
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Volume Title
Publisher
Humana Press Inc.
Abstract
Cell-free protein synthesis offers fast and inexpensive access to selectively isotope labeled proteins that can be measured by NMR spectroscopy in the presence of all the unlabeled proteins in the reaction mixture. No chromatographic purification is required. Using an extract from Escherichia coli in a simple dialysis system, the target protein can be prepared at a typical concentration of about 1 mg/ml, which is sufficient for subsequent analysis by NMR. This chapter describes in detail the protocol used in the authors' laboratory.
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Keywords
Keywords: Escherichia coli protein; nitrogen; article; cell free system; chemistry; Escherichia coli; genetics; isotope labeling; metabolism; methodology; nuclear magnetic resonance spectroscopy; protein synthesis; Cell-Free System; Escherichia coli; Escherichia co
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Type
Book chapter
Book Title
Structural Proteomics: High Throughput Methods
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Restricted until
2037-12-31
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