Cell-free protein synthesis for analysis by NMR spectroscopy

Date

2008

Authors

Apponyi, Margit
Ozawa, Kiyoshi
Dixon, Nicholas
Otting, Gottfried

Journal Title

Journal ISSN

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.

Description

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

Citation

Source

Type

Book chapter

Book Title

Structural Proteomics: High Throughput Methods

Entity type

Access Statement

License Rights

DOI

Restricted until

2037-12-31