Creation and X-ray Structure Analysis of the Tumor Necrosis Factor Receptor-1-selective Mutant of a Tumor Necrosis Factor-α Antagonist

Shibata, Hiroko; Yoshioka, Yasuo; Ohkawa, Akiko; Minowa, Kyoko; Mukai, Yohei; Abe, Yasuhiro; Taniai, Madoka; Nomura, Tetsuya; Kayamuro, Hiroyuki; Nabeshi, Hiromi; Sugita, Takahisa; Imai, Shunji; Nagano, Kazuya; Yoshikawa, Tomoaki; Fujita, Takuya; Nakagawa, Shinsaku; Yamamoto, Akira; Ohta, Toshio; Hayakawa, Tetsuo; Mayumi, Tadanori; Vandenabeele, Peter; Aggarwal, Bharat B.; Nakamura, Teruya; Yamagata, Yuriko; Tsunoda, Shin-ichi; Kamada, Haruhiko; Tsutsumi, Yasuo

doi:10.1074/jbc.m707933200

Cited by 91 publications

(64 citation statements)

References 50 publications

(51 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Rational design approaches have been previously used to design receptor-selective TNF-␣ variants (35) and TRAIL variants (22,23,26,36). Directed evolution methods employing phage display were also used to generate receptorselective TRAIL variants (37), a receptor-selective TNF␣ antagonist (38), and a LIGHT/LT␤ variant that does not bind DcR3 (39). In addition, selective receptor activation can also be achieved by the use of receptor-specific agonistic antibodies (40,41) or other binding scaffolds (42).…”

Section: Discussionmentioning

confidence: 99%

The Design and Characterization of Receptor-selective APRIL Variants

Kimberley¹,

Sloot

Guadagnoli³

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

Background: APRIL binds two receptors, BCMA and TACI, so separating signaling outcomes is difficult. Results: We used an algorithm to design a variant of APRIL that specifically binds BCMA and two variants that selectively bind TACI. Conclusion: TACI and BCMA signals differ in the context of B cell stimulation. Significance: These variants will help decipher APRIL signaling in physiology and disease settings.

show abstract

Section: Discussionmentioning

confidence: 99%

The Design and Characterization of Receptor-selective APRIL Variants

Kimberley¹,

Sloot

Guadagnoli³

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…It is known that molecular modifications of proteins such as PEGylation are effective in suppressing immunogenicity, degradation by proteases, and excretion into the urine (24,29). We previously reported that N terminus-specific monoPEGylation of R1antTNF improved in vivo stability (20,21). However, because R1antTNF adopts a homotrimeric structure composed of three monomers, there are three amino groups arising from each N-terminal residue that may be subject to modification by PEG (Fig.…”

Section: Molecular Modification Efficiency Of Scr1anttnfmentioning

confidence: 99%

“…Therefore, to increase the half-life of the protein, we have applied site-specific PEGylation for R1antTNF, which is a modification technique using polyethylene glycol (PEG) developed in our previous work (23)(24)(25). PEGylation significantly improves the in vivo stability and therapeutic effect of R1antTNF without any associated loss of bioactivity (20,21). However, given that R1antTNF exists as a trimeric structure, there are problems such as a reduction in biological activity caused by intermolecular dissociation.…”

mentioning

confidence: 99%

See 1 more Smart Citation

A trimeric structural fusion of an antagonistic tumor necrosis factor-α mutant enhances molecular stability and enables facile modification

Inoue

Ando

Kamada

et al. 2017

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Edited by Peter CresswellTumor necrosis factor-␣ (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 in vitro and in vivo. 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 in vitro molecular properties of receptor selectivity and antagonistic activity but displayed a marked increase in thermal stability. The residence time of scR1antTNF in vivo 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.Recently, TNF inhibitors in various molecular formats such as a neutralizing antibody (infliximab or adalimumab), soluble receptor (etanercept), or a PEGylated antibody Fab fragment (certolizumab pegol), have been clinically used as anti-TNF drugs to treat autoimmune diseases such as rheumatoid arthritis, multiple sclerosis, and ulcerative colitis (1-3). These drugs elicit a highly beneficial therapeutic effect, but this treatment may lead to serious complications including bacterial or viral infection (4, 5), demyelination (6), and lupus-like syndrome (7). Therefore, biopharmaceutical development of this treatment with a new modality is sought.TNF binds to two receptor subtypes to exert its biological functions: TNFR1 3 and TNFR2 (8). TNFR1 is expressed in diverse cell types and raises the inflammatory response, whereas TNFR2, which is limited to expression in immune cells, endothelial cells, and neuronal cells, induces cell survival and proliferation. For example, it has been reported that the incidence and severity of arthritis were lower and milder in TNFR1 knock-out mice than in wild-type mice (9). Moreover, TNFR1 knock-out mice or TNFR1/TNFR2 double knock-out mice display less severe symptoms of experimental autoimmune encephalomyelitis symptoms than wild-type mice (10). Previous studies demonstrated that transgenic mice overexpressing human TNF develop severe arthritis (11, 12) or demyelination (13). Thus, the involvement of TNFR1 in autoimmune diseases is strongly implicated as human TNF is unable to signal

show abstract

“…This molecular crosstalk makes it complicated to elucidate the roles of each TNF receptor in cell proliferation and apoptosis. In order to activate TNFR2 specifically, a TNFR2-specific agonist (R2-7) was generated from a mutated TNF library using phage-display technology (Shibata et al, 2008;Nomura et al, 2009). By using this technology, R2-7 was identified as a TNFR2-selective TNF mutant protein with six amino acid mutations (positions at 29, 31, 32, 145, 146 and 147) from wild-type TNF (wtTNF).…”

Section: Introductionmentioning

confidence: 99%

Aminopeptidase P3 (APP3), a novel member of the TNF/TNFR2 signaling complex, induces phosphorylation of JNK

Inoue

Kamada

Abe

et al. 2015

Journal of Cell Science

Self Cite

View full text Add to dashboard Cite

Tumor necrosis factor (TNF) is an important mediator that triggers onset of autoimmune diseases and exerts its biological effects by interacting through two types of receptors, TNFR1 and TNFR2. The TNFR2 signaling has significant potential to exert pro-survival and protective roles in several disorders. Unlike TNFR1 signaling, however, the mechanism of TNFR2 signal transduction is poorly understood, and few of its adapter molecules are known. The present study utilized a proteomics approach to search for adapter molecules in the TNFR2 signaling complex and identified aminopeptidase P3 (APP3) to be a key molecule. One of its two isoforms, mitochondrial APP3 (APP3m) but not cytosolic APP3 (APP3c), was recruited to TNFR2 and shown to regulate TNF/TNFR2-dependent JNK phosphorylation. Furthermore, APP3m was released from mitochondria upon TNF stimulation in the absence of mitochondrial outer membrane permeabilization (MOMP). The observation of increased cell death by down-regulation of APP3m also suggested that APP3m exerts an anti-apoptotic function. These findings reveal that APP3m is a new member of the TNF/TNFR2 signaling complex and characterize an APP3-mediated TNFR2 signal transduction mechanism that induces JNK activation.

show abstract

Creation and X-ray Structure Analysis of the Tumor Necrosis Factor Receptor-1-selective Mutant of a Tumor Necrosis Factor-α Antagonist

Cited by 91 publications

References 50 publications

The Design and Characterization of Receptor-selective APRIL Variants

The Design and Characterization of Receptor-selective APRIL Variants

A trimeric structural fusion of an antagonistic tumor necrosis factor-α mutant enhances molecular stability and enables facile modification

Aminopeptidase P3 (APP3), a novel member of the TNF/TNFR2 signaling complex, induces phosphorylation of JNK

Contact Info

Product

Resources

About