Skip Navigation



Journal of Biochemistry Advance Access published online on April 22, 2009
Journal of Biochemistry, doi:10.1093/jb/mvp065
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow All Versions of this Article:
146/2/167    most recent
mvp065v1
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 Mukai, Y.
Right arrow Articles by Tsutsumi, Y.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mukai, Y.
Right arrow Articles by Tsutsumi, Y.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The authors 2009. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

Fast binding kinetics and conserved 3D structure underlie the antagonistic activity of mutant TNF: useful information for designing artificial proteo-antagonists.

Yohei Mukai1,2, Teruya Nakamura3, Yasuo Yoshioka2,4, Hiroko Shibata2, Yasuhiro Abe2, Tetsuya Nomura1,2, Madoka Taniai5, Tsunetaka Ohta5, Shinsaku Nakagawa1, Shin-ichi Tsunoda2, Haruhiko Kamada2, Yuriko Yamagata3 and Yasuo Tsutsumi1,2,*

1. Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan
2. Laboratory of Pharmaceutical Proteomics, National Institute of Biomedical Innovation (NiBio), Osaka 567-0085, Japan
3. Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan
4. The Center for Advanced Medical Engineering and Informatics, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan
5. Hayashibara Biochemical Laboratories, Inc., 1-2-3 Shimoishii, Okayama 702-8006, Japan

* Corresponding Author Yasuo Tsutsumi, PhD Department of Toxicology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871, Japan. Tel: +81-6-6879-8230, Fax: +81-6-6879-8234, E-mail address: ytsutsumi{at}phs.osaka-u.ac.jp

Received February 18, 2009; Accepted March 17, 2009


  Abstract

Tumor necrosis factor (TNF) is an important cytokine that induces an inflammatory response predominantly through the TNF receptor-1 (TNFR1). A crucial strategy for the treatment of many autoimmune diseases, therefore, is to block the binding of TNF to TNFR1. We previously identified a TNFR1-selective antagonistic mutant TNF (R1antTNF) from a phage library containing 6 randomized amino acid residues at the receptor binding site (amino acids 84-89). Two R1antTNFs, R1antTNF-T2 (A84S, V85T, S86T, Y87H, Q88N, and T89Q) and R1antTNF-T8 (A84T, V85P, S86A, Y87I, Q88N, and T89R), were successfully isolated from this library.

Here, we analyzed R1antTNF-T8 using surface plasmon resonance spectroscopy and X-ray crystallography to determine the mechanism underlying the antagonistic activity of R1antTNF. The kinetic association/dissociation parameters of R1antTNF-T8 were higher than those of wild-type TNF, indicating more rapid bond dissociation. X-ray crystallographic analysis suggested that the binding mode of the T89R mutation changed from a hydrophobic to an electrostatic interaction, which may be responsible for the antagonistic behavior of R1antTNF. Knowledge of these structure-function relationships will facilitate the design of novel TNF inhibitors based on cytokine structure.

Key Words: Antagonistic activity, Mutant, Tumor necrosis factor (TNF), X-ray crystallography, Structure-function relationship


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




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.