Springer Protocols: Abstract: Production of Protein for Nuclear Magnetic Resonance Study Using the Wheat Germ Cell-Free System

Production of Protein for Nuclear Magnetic Resonance Study Using the Wheat Germ Cell-Free System

By: Toshiyuki Kohno², Yaeta Endo³

Abstract

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Nuclear magnetic resonance (NMR) methods have been developed to determine the three-dimensional structures of proteins, to estimate protein folding, and to discover high-affinity ligands for proteins. However, one of the difficulties encountered in the application of such NMR methods to proteins is that we should obtain milligram quantities of ¹⁵N and/or ¹³C-labeled pure proteins of interest.

Here, we describe the method to produce proteins for NMR experiments using the improved wheat germ cell-free system, which exhibits several attractive features for high-throughput NMR study of proteins.

Affiliation(s): (2) Molecular Structure Research Group, Mitsubishi Kagaku Institute of Life Sciences (MITILS), Tokyo, Japan
(3) Cell-Free Science and Technology Research Center, Ehime University, Matsuyama, Japan

Book Title: In Vitro Transcription and Translation Protocols

Series: Methods in Molecular Biology | Volume: 375 | Pub. Date: May-03-2007 | Page Range: 257-272 | DOI: 10.1007/978-1-59745-388-2_13

Subject: Genetics/Genomics

Key Words: Wheat germ - cell-free - protein synthesis - NMR - HSQC - protein folding - transaminase - glutamine synthase - selective labeling

References

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