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Structure-based design of an agonistic peptide targeting Fas

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Abstract

A small agonistic peptide FRAP-4 (WEWT, Fas reactive peptide-4) that binds to the human Fas molecule was discovered using our computer screening strategy named the Amino acid Complement Wave (ACW) method, which is based on the complementarities of interacting amino acids between comprehensive testing peptides and a target protein surface pocket. In silico docking studies demonstrated the specific interaction of FRAP-4 with the main Fas ligand (FasL) binding domain in the Fas molecule. An octamer of this peptide produced by carboxyl terminal linkages of polylysine branches (MAP), (FRAP-4)8-MAP, effectively induced apoptosis in human ovarian cancer cell line NOS4 cells that was associated with the activation of caspases-8, -9 and -3, and the cleavage of PARP. Alanine substitution of the N-terminal W in FRAP-4 resulted in complete loss of FasL-mimetic action of (FRAP-4)8-MAP, suggesting that the aromatic functionality at the N-terminal position W appears to play an essentially important role in Fas binding ability. These observations indicate that the FasL-mimetic peptide should serve as an excellent starting point for the design of effective compounds with FasL-mimetic activity. Furthermore, the ACW method for the structure-based design of optimized small peptides against receptor molecules such as Fas could open new avenues for the development of peptide mimetic and nonpeptidic organic forms to generate novel effective pharmaceuticals.

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Abbreviations

ACW:

amino acid complement wave

BPB:

bromophenol blue

BSA:

bovine serum albumin

CAD:

caspase-activated DNase

3D:

three-dimensional

DTT:

dithiotheritol

FRAP:

Fas reactive peptide

IAP:

inhibitor of apoptosis protein

PI:

propidum iodide

mAb:

monoclonal antibody

pAb:

polyclonal antibody

PVDF:

poly(vinylidene fluoride)

RBD:

receptor-binding domain

SBDD:

structure-based drug design

SDS:

sodium dodecyl sulfate

PAGE:

polyacrylamide gel electrophoresis

TNFR:

tumor necrosis factor receptor

TRAILR:

TNF-related apoptosis-inducing ligand receptor.

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Authors and Affiliations

  1. Research Institute for Biological Sciences, Tokyo University of Science, 2669 Yamazaki Noda, Chiba, 278-0022, Japan

    A. Yoshimori & S. Tanuma

  2. Genome and Drug Research Center, Tokyo University of Science, 2641 Yamazaki Noda, Chiba, 278-8510, Japan

    R. Takasawa & S. Tanuma

  3. Department of Equipment Center for Research and Education, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan

    A. Hayakawa

  4. Department of Molecular Neurosurgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan

    M. Mizuno

  5. Department of Neurosurgery, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, 466-8550, Japan

    J. Yoshida

  6. Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki Noda, Chiba, 278-8510, Japan

    S. Tanuma

  7. Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki Noda, Chiba, 278-8510, Japan

    S. Tanuma

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  1. A. Yoshimori
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  2. R. Takasawa
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  3. A. Hayakawa
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  4. M. Mizuno
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  5. J. Yoshida
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  6. S. Tanuma
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Correspondence to S. Tanuma.

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Yoshimori, A., Takasawa, R., Hayakawa, A. et al. Structure-based design of an agonistic peptide targeting Fas. Apoptosis 10, 323–329 (2005). https://doi.org/10.1007/s10495-005-0806-6

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