Papers - USUI Kenji
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Protein-detection microarrays using structure-based designed peptide libraries Reviewed
Kenji Usui, Tetsunori Ojima, Masato Suzuki, Sin-ya Watanabe, Kin-ya Tomizaki, Kiyoshi Nokihara, Hisakazu Mihara
Peptides 2004, Proceedings 401 - 402 2005
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Peptide Arrays with Designed Secondary Structures for Protein Characterization Using Fluorescent Fingerprint Patterns. Reviewed
臼井健二
Biopolymers ( 76 ) 129 - 139 2004.11
Single Work
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Peptide arrays with designed alpha-helical structures for characterization of proteins from FRET fingerprint patterns Reviewed
Kenji Usui, Mizuki Takahashi, Kiyoshi Nokihara, Hisakazu Mihara
MOLECULAR DIVERSITY 8 ( 3 ) 209 - 218 2004.9
Publisher：SPRINGER
A practical high-throughput protein detection system is described, based on synthetic peptide arrays consisting of designed alpha-helical peptides, detected by fluorescence resonance energy transfer (FRET). Initially a model alpha-helical peptide known to interact with a structured protein, calmodulin, was selected to establish the strategy for high-throughput detection. In comparison to peptides with a single probe, a much higher FRET response has been observed with two fluorescent probes (7-diethylaminocoumarin-3-carboxylic acid and 5(6)-carboxy-fluorescein) at both termini of the synthetic peptides. To establish a reproducible high-throughput detection system, peptides were also immobilized onto a solid surface for detection of the target proteins. A small library of 112 different peptides was constructed, based on a model of the alpha-helical peptide with systematic replacement of residues carrying specific charges and/or hydrophobicities. The library was used to effectively characterize various proteins, giving their own 'protein fingerprint' patterns. The resulting 'protein fingerprints' correlate with the recognition properties of the proteins. The present microarray with designed synthetic peptides as the capturing agents is promising for the development of protein detection chips.
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Development of a practical protein-chip using designed synthetic peptide-arrays Reviewed
K Nokihara, T Ohyama, K Usui, K Yonemura, K Tomizaki, H Mihara
KOBUNSHI RONBUNSHU 61 ( 10 ) 523 - 532 2004
Publisher：SOC POLYMER SCIENCE JAPAN
Novel high-throughput technologies, which can replace conventional electrophoresis and mass spectroscopic analyses, are in great demand for understanding the structure-function relationships of proteins. For a practical protein-detection system, arrays with immobilized designed synthetic peptides have been constructed. Peptides were labeled with fluorescent dyes in order to achieve high sensitivity. Two different types of prototype-arrayer have been constructed: one has a micro-dispensing system and the other a spot-printing system. Combinatorial peptide libraries, which consisted of beta-loop, beta-strand and alpha-helical peptides, were constructed by improved highly efficient solid-phase synthesis. Labeled peptides were covalently immobilized on solid supports such as precision glass plates. Various proteins were characterized with these peptide-arrays using a fluorescent scanner to give the "protein fingerprint" which is characteristic of the individual proteins. The results of the present prototype system demonstrate the practicality of protein-chips as a new generation of biochips.
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Development of Peptide-Chips Focusing on a High Throughput Protein-Detection System Reviewed
Kiyoshi Nokihara, Takafumi Ohyama, Kenji Usui, Koichi. Yoneyama, Mizuki Takahashi, Hisakazu Mihara
Innovation and Perspectives in Solid Phase Synthesis & Combinatorial Libraries 83 - 88 2004
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Peptide arrays with designed secondary structures for protein characterization using fluorescent fingerprint patterns Reviewed
K Usui, T Ojima, M Takahashi, K Nokihara, H Mihara
BIOPOLYMERS 76 ( 2 ) 129 - 139 2004
Publisher：JOHN WILEY & SONS INC
To realize a practical high-throughput protein-detection system, novel peptide arrays have been constructed using designed peptide libraries with loop, a-helix, or P-strand structures. Here, we describe the overview of the reported deigned peptide arrays with loop and a-helix structures and the new results of those with beta-strand structures. Initially, several model peptides known to interact with model structured proteins were selected to establish the present strategy for high-throughput detection of proteins. The fluorescent probes and suitable scaffolds of peptides were examined for the effective detection of proteins. The detection methods were established in solution and in an immobilized manner using the model systems. In the case of a-helix peptide, the response of a peptide with fluorescent resonance energy tran, fer between two probes at both termini was several times higher than that of a peptide with a single probe. In the cases of peptides with other structures, however, proteins were effectively detectable even by the fluorescent change of one probe. Furthermore, structurally focused libraries consisting of a total of ca. 250 different peptides based on the model pcptides with secondary and/or tertiary structures were constructed with systematic replacement of residues. Using these libraries, various proteins were characterized effectively to give their own fluorescent "protein fingerprint" patterns. The resulting protein fingerprints correlated with the recognition properties of the proteins. These studies demonstrate that arrays with peptide libraries based on designed structures can be promising tools for detecting the target proteins. Designed synthetic peptides play roles as the capturing agents to be developed for practical protein chips. (C) 2004 Wiley Periodicals, Inc.
DOI： 10.1002/bip.10568
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Peptide microarrays using structure-based peptide libraries for protein chips Reviewed
Kenji Usui, Mizuki Takahashi, Tetsunori Ojima, Masato Suzuki, Kiyoshi Nokihara, Eiichi Tamiya, Hisakazu Mihara
Peptide Revolution: Genomics, Proteomics & Therapeutics 258 - 259 2004