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Structural Conversion Rate Changes of Recombinant Bovine Prion by Designed Synthetic Peptides

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Abstract

An understanding of structural changes and self-assembly of proteins, which are thought to involve specific peptide–peptide interactions, will contribute to the development of therapeutic agents and diagnosis for the detection of conformational diseases. We hypothesize that certain peptides may contribute to the conformational change of prion proteins. The present paper describes the discovery of prion-related synthetic peptides which influence structural conversion of recombinant bovine prion protein. The peptides designed are prion-protein fragments containing core domains consisting of α-helical (human prion protein fragment 180–195) and known β-sheet (human prion protein fragment 169–175) structures. Additionally several reported known β-sheet breaker peptides and a conjugate consisting of β-sheet and α-helix segments based on the secondary structures of human prion protein, designated HPPSH, have been chemically synthesized by the conventional Fmoc solid-phase method and characterized by circular dichroism and the Thioflavin T fluorescence method. Our data indicated that the co-existence of peptides, HPPSH or other prion fragment peptides involving toxic core sequence (the fragment 106–126), influenced the kinetic rate of aggregation and the lag-time of fibril formation of recombinant bovine prion protein except the core sequence itself. The method will be used for discovery of responsible material from natural resources. And designed peptides can be also used for bio-detection.

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Abbreviations

Fmoc:

9-Fluorenylmethoxycarbonyl

HBTU:

(2-(1H-Benzotriazole-1-yl)-1,1,3,3-tetramethylaminiumhexafluoro-phosphate

HOBt:

1-Hydroxybenzotriazol

DIEA:

N,N-Diisopropyl ethylamine

TFA:

Trifluoroacetic acid

DMF:

N,N-Dimethylformamide

ThT:

Thioflavin T

CD:

Circular dichroism

IR:

Infrared ray

TEM:

Transmission electron microscopy

TFE:

Trifluoroethanol

hPrP:

Human prion protein

rbPrP:

Recombinant bovine prion protein

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Acknowledgments

The authors thank Mr. K. Fukano, Tokyo University of Agriculture, Ms. M. Miyajima, HiPep Laboratories for their technical assistance, Drs. S. Mohri, T. Yokoyama and K. Kasai, Prion Disease Research Center, National Institute of Animal Health, Tsukuba, Japan for discussion and Dr. V. Wray, Helmholtz Centre for Infection Research, Braunschweig, Germany, for variable discussion with linguistic advice. A part of this work was funded by the Research and Development Program for New Bio-industry Initiatives, National Agriculture and Food Research Organization.

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Authors and Affiliations

  1. HiPep Laboratories, Nakatsukasa-cho 486-46, Kamigyo-ku Kyoto, 602-8158, Japan

    Akiyoshi Hirata & Kiyoshi Nokihara

  2. Tokyo University of Agriculture, Tokyo, 156-0054, Japan

    Shunsuke Yajima & Tadashi Yasuhara

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  1. Akiyoshi Hirata
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  2. Shunsuke Yajima
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  3. Tadashi Yasuhara
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  4. Kiyoshi Nokihara
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Correspondence to Kiyoshi Nokihara.

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Hirata, A., Yajima, S., Yasuhara, T. et al. Structural Conversion Rate Changes of Recombinant Bovine Prion by Designed Synthetic Peptides. Int J Pept Res Ther 18, 217–225 (2012). https://doi.org/10.1007/s10989-012-9294-z

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