Journal Article

A Novel Way to Express Proline-Selectively Labeled Proteins with a Wheat Germ Cell–Free Protein Synthesis System

Masato Shimizu, Takahisa Ikegami, Koichi Akiyama and Eugene Hayato Morita

in The Journal of Biochemistry

Published on behalf of The Japanese Biochemical Society

Volume 140, issue 3, pages 453-456
Published in print September 2006 | ISSN: 0021-924X
Published online September 2006 | e-ISSN: 1756-2651 | DOI: http://dx.doi.org/10.1093/jb/mvj174
A Novel Way to Express Proline-Selectively Labeled Proteins with a Wheat Germ Cell–Free Protein Synthesis System

Show Summary Details

Preview

For high-throughput protein structural analyses, it is essential to develop a reliable protein overexpression system. Although many protein overexpression systems, such as ones involving Escherichia coli cells, have been developed, the number of overexpressed proteins exhibiting the same biological activities as those of the native ones is limited. A novel wheat germ cell–free protein synthesis system was developed recently, and most of the synthesized proteins that should function in solution were found to be in soluble forms. This suggests the applicability of this protein synthesis method to determination of the functional structures of soluble proteins. In our previous work, we developed a selective labeling technique for amino acids having amide functional groups (other than proline residues) involving the use of several inhibitors for transaminases. This paper in turn describes a proline-selective labeling technique. Based on our results, we have succeeded in constructing a complete amino acid selective labeling technique for the wheat germ cell–free protein synthesis system.

Keywords: cell-free; HNCO; proline; selective labeling; wheat germ

Journal Article.  2797 words.  Illustrated.

Subjects: Biochemistry

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content.