A Trimeric Structural Fusion of an Antagonistic Tumor Necrosis Factor-α Mutant Enhances Molecular Stability and Enables Facile Modification

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2017 Feb 26

PMID 28235800

Citations 8

Authors

Masaki Inoue

Daisuke Ando

Haruhiko Kamada

Shintaro Taki

Mayumi Niiyama

Yohei Mukai

Takashi Tadokoro

Katsumi Maenaka

Taisuke Nakayama

Yuji Kado

Tsuyoshi Inoue

Yasuo Tsutsumi

Shin-Ichi Tsunoda

Affiliations

Soon will be listed here.

Abstract

Tumor necrosis factor-α (TNF) exerts its biological effect through two types of receptors, p55 TNF receptor (TNFR1) and p75 TNF receptor (TNFR2). An inflammatory response is known to be induced mainly by TNFR1, whereas an anti-inflammatory reaction is thought to be mediated by TNFR2 in some autoimmune diseases. We have been investigating the use of an antagonistic TNF mutant (TNFR1-selective antagonistic TNF mutant (R1antTNF)) to reveal the pharmacological effect of TNFR1-selective inhibition as a new therapeutic modality. Here, we aimed to further improve and optimize the activity and behavior of this mutant protein both and Specifically, we examined a trimeric structural fusion of R1antTNF, formed via the introduction of short peptide linkers, as a strategy to enhance bioactivity and molecular stability. By comparative analysis with R1antTNF, the trimeric fusion, referred to as single-chain R1antTNF (scR1antTNF), was found to retain molecular properties of receptor selectivity and antagonistic activity but displayed a marked increase in thermal stability. The residence time of scR1antTNF was also significantly prolonged. Furthermore, molecular modification using polyethylene glycol (PEG) was easily controlled by limiting the number of reactive sites. Taken together, our findings show that scR1antTNF displays enhanced molecular stability while maintaining biological activity compared with R1antTNF.

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