Peptide arrays with designed alpha-helical structures for characterization of proteins from FRET fingerprint patterns

Molecular Diversity
Kenji UsuiHisakazu Mihara

Abstract

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 agen...Continue Reading

Citations

Aug 2, 2006·Molecular BioSystems·Sinya WatanabeHisakazu Mihara
Oct 12, 2010·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Ákos I FábiánAttila Jenei
Dec 1, 2010·Pharmaceutical Research·Luis BerradeJulio A Camarero
Mar 26, 2005·Chembiochem : a European Journal of Chemical Biology·Kin-ya TomizakiHisakazu Mihara
Jun 16, 2006·Journal of Molecular Recognition : JMR·Mark R ContarinoIrwin Chaiken
Jun 12, 2013·Chemical Communications : Chem Comm·Kenji UsuiHisakazu Mihara
Jan 1, 2015·Microarrays·Amir SyahirHisakazu Mihara
Dec 13, 2006·Molecular BioSystems·Kenji UsuiHisakazu Mihara
May 14, 2009·Accounts of Chemical Research·Mahesh UttamchandaniShao Q Yao
Oct 13, 2006·Bioorganic & Medicinal Chemistry Letters·Kenji UsuiHisakazu Mihara
Jan 18, 2007·Journal of the American Chemical Society·Söhnke VossRoland Brock

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