A novel peptide microarray for protein detection and analysis utilizing a dry peptide array system.

A novel dry peptide microarray system has been constructed that affords a practical solution for protein detection and analysis. This system is an array preparation and assay procedure under dry conditions that uses designed peptides as non-immobilized capture agents for the detection of proteins. The system has several advantages that include its portability and ease-of-use, as well as the fact that vaporization of sample solutions need not be considered. In this study, various proteins have been characterized with an alpha-helical peptide mini-library. When proteins were added to the peptide library array, the fluorescent peptides showed different fluorescent intensities depending on their sequences. The patterns of these responses could be regarded as 'protein fingerprints' (PFPs), which are sufficient to establish the identities of the target proteins. Furthermore, statistical analysis of the resulting PFPs was performed using cluster analysis. The PFPs of the proteins were clustered successfully depending on their families and binding properties. Additionally, the target protein was characterized using a nanolitre system and could be detected down to 1.2 fmol. These studies imply that the dry peptide array system is a promising tool for detecting and analyzing target proteins. The dry peptide array will play a role in development of high-throughput protein-detecting nano/micro arrays for proteomics and ligand screening studies.

[1]  Dieter Stoll,et al.  Protein microarray technology. , 2002, Frontiers in bioscience : a journal and virtual library.

[2]  P. Brown,et al.  Exploring the metabolic and genetic control of gene expression on a genomic scale. , 1997, Science.

[3]  Kenji Usui,et al.  Protein‐Detecting Microarrays: Current Accomplishments and Requirements , 2005, Chembiochem : a European journal of chemical biology.

[4]  M. Lesaicherre,et al.  Enzymatic profiling system in a small-molecule microarray. , 2003, Organic letters.

[5]  Kiyoshi Nokihara,et al.  Construction of a protein-detection system using a loop peptide library with a fluorescence label. , 2003, Chemistry & biology.

[6]  W. Chan,et al.  Fmoc Solid Phase Peptide Synthesis: A Practical Approach (Practical Approach Series) , 2019 .

[7]  J. Cox,et al.  Structural changes in melittin and calmodulin upon complex formation and their modulation by calcium. , 1983, Biochemistry.

[8]  N. Anderson,et al.  Proteome and proteomics: New technologies, new concepts, and new words , 1998, Electrophoresis.

[9]  Kenji Usui,et al.  Peptide arrays with designed α-helical structures for characterization of proteins from FRET fingerprint patterns , 2004, Molecular Diversity.

[10]  C. Cantor,et al.  Biophysical Chemistry: Part II: Techniques for the Study of Biological Structure and Function , 1980 .

[11]  T. Kodadek Protein microarrays: prospects and problems. , 2001, Chemistry & biology.

[12]  S. Schreiber,et al.  Printing proteins as microarrays for high-throughput function determination. , 2000, Science.

[13]  A. Podtelejnikov,et al.  Linking genome and proteome by mass spectrometry: large-scale identification of yeast proteins from two dimensional gels. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Dev Kambhampati,et al.  Protein microarray technology , 2003 .

[15]  A. Mirzabekov,et al.  Protein microchips: use for immunoassay and enzymatic reactions. , 2000, Analytical biochemistry.

[16]  Anomalous reflection of gold applicable for a practical protein-detecting chip platform. , 2005, Molecular bioSystems.

[17]  G. Fasman,et al.  Computed circular dichroism spectra for the evaluation of protein conformation. , 1969, Biochemistry.

[18]  W. DeGrado,et al.  The interaction of calmodulin with amphiphilic peptides. , 1985, The Journal of biological chemistry.

[19]  J. Reymond,et al.  Enzyme activity fingerprinting with substrate cocktails. , 2004, Journal of the American Chemical Society.

[20]  S. Gygi,et al.  Correlation between Protein and mRNA Abundance in Yeast , 1999, Molecular and Cellular Biology.

[21]  K. Usui,et al.  A designed glycopeptide array for characterization of sugar-binding proteins toward a glycopeptide chip technology , 2005 .

[22]  S. Gygi,et al.  Quantitative analysis of complex protein mixtures using isotope-coded affinity tags , 1999, Nature Biotechnology.

[23]  H. Mantsch,et al.  Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment. , 1993, Biochemistry.

[24]  F. Toda,et al.  Inclusion Complexes and Guest-Induced Color Changes of pH-Indicator-Modified .beta.-Cyclodextrins , 1994 .

[25]  M. Snyder,et al.  Protein arrays and microarrays. , 2001, Current opinion in chemical biology.

[26]  I. Hamachi,et al.  Semi-wet peptide/protein array using supramolecular hydrogel , 2004, Nature materials.

[27]  S. Nock,et al.  Recent developments in protein microarray technology. , 2003, Angewandte Chemie.

[28]  William F. DeGrado,et al.  How calmodulin binds its targets: sequence independent recognition of amphiphilic α-helices , 1990 .

[29]  P. Mitchell A perspective on protein microarrays , 2002, Nature Biotechnology.

[30]  Jean‐Louis Reymond,et al.  Classifying Enzymes from Selectivity Fingerprints , 2004, Chembiochem : a European journal of chemical biology.

[31]  Jean-Louis Reymond,et al.  Enzyme fingerprints of activity, and stereo- and enantioselectivity from fluorogenic and chromogenic substrate arrays. , 2002, Chemistry.

[32]  T Isobe,et al.  BIA-MS-MS: biomolecular interaction analysis for functional proteomics. , 2001, Trends in biotechnology.

[33]  Ronald Frank,et al.  Multiplexed sorting of libraries on libraries: A novel method for empirical protein design by affinity-driven phage enrichment on synthetic peptide arrays , 2004, Molecular Diversity.

[34]  Kenji Usui,et al.  Peptide arrays with designed secondary structures for protein characterization using fluorescent fingerprint patterns. , 2004, Biopolymers.