Skip Navigation

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Nishi, N.
Right arrow Articles by Ishida, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Nishi, N.
Right arrow Articles by Ishida, T.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

J. Biochem, 1998, Vol. 123, No. 1 157-161
© 1998 Japanese Biochemical Society

research-article

Expression of a Synthetic Gene for Initiation Factor 4E-Binding Protein 1 in Escherichia coli and Its Interaction with eIF-4E and eIF-4E-m7GTP Complex1

Noriko Nishi, Shigenobu Morino, Koji Tomoo, Takeshi Youtani and Toshimasa Ishida2

Department of Physical Chemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara Takatsuki, Osaka 569-11

2 To whom correspondence should be addressed. Tel/Fax: +81-726-90-1068, E-mail: ishida{at}oysun01.oups.ac.jp

An artificial gene coding for the human initiation factor (elF) 4E-binding protein 1 (4E-BP1) was chemically synthesized and cloned. Although the expression of the 4E-BP1 gene alone has not yet been accomplished, the gene was expressed in Escherichia coli [BL21(DE3)] as a fusion gene with the glutathione-S-transferase (GST) gene using a prokaryotic gene fusion vector (pGEX-4T-2), which contains a gene sequence coding the cleavage site for a specific protease, {alpha}-thrombin. The fusion gene product was purified to homogeneity by glutathione Sepharose-4B affinity column chromatography. It was shown by m7GTP- and glutathione-affinity chromatography that the binding ability of 4E-BP1 to eIF-4E is nearly the same as that to the eIF-4E-m7GTP complex, implying different binding sites of eIF-4E and its nonallosteric obligation for 4E-BP1 and mRNA cap structure. In contrast with the binding of eIF-4E to the mRNA cap structure, where some functional amino acids play an important role in the binding, the binding to 4E-BP1 was suggested to occur via multiple nonspecific interactions.

1 This study was supported in part by The Science Research Promotion Fund from Japan Private School Promotion Foundation.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Mol. Cell. Biol.Home page
A. J. Waskiewicz, J. C. Johnson, B. Penn, M. Mahalingam, S. R. Kimball, and J. A. Cooper
Phosphorylation of the Cap-Binding Protein Eukaryotic Translation Initiation Factor 4E by Protein Kinase Mnk1 In Vivo
Mol. Cell. Biol., March 1, 1999; 19(3): 1871 - 1880.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.