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Interactions between peptides containing nucleobase amino acids and T7 phages displaying S. cerevisiae proteins
Author(s) -
Watanabe Sinya,
Tomizaki Kinya,
Takahashi Tsuyoshi,
Usui Kenji,
Kajikawa Kotaro,
Mihara Hisakazu
Publication year - 2007
Publication title -
peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.20662
Subject(s) - proteome , biochemistry , chemistry , phage display , nucleic acid , amino acid , peptide , ribosomal protein , computational biology , biology , gene , rna , ribosome
The importance of high‐throughput analyses of protein abundances and functions is interestingly increasing in genomic/proteomic studies. In such postgenome sequencing era, a protein‐detecting chip, in which a large number of molecules specifically capturing target proteins (capturing agents) such as antibodies, recombinant proteins, and small molecules are arrayed onto solid, wet, or semi‐wet substrates, enables comprehensive analysis of proteomes by a single experiment. However, whole proteomes are generally complicated for comprehensive analyses so that alternative approaches to subproteome analysis categorized by protein functions and binding properties (focused proteome) would be effective. Approaching the goal of development of designed peptide chip for protein analysis, diversity increases in peptide structures and validation of target proteins are needed. We herein describe design and synthesis of nucleobase amino acid (NBA)‐containing peptides, selection of nucleic acid‐related proteins derived from S. cerevisiae , and detection of interactions between NBA‐containing peptides and T7 phages displaying proteins by both enzyme‐linked immunosorbent assays (ELISA) and label‐free anomalous reflection of gold (AR) measurements. Twenty‐eight phage clones were obtained by the phage‐display method and sequenced. Ten of 28 clones were expected to be nucleic acid‐related proteins including initiation factor, TYB protein, ribosomal proteins, elongation factor, ATP synthase subunit, GTP‐binding protein, and ribonuclease. Other phage clones encoded several classes of enzymes such as reductase, oxidase, aldolase, metalloprotease, and hexokinase. Both ELISA and AR measurements suggested that the methodology of in vitro selection for recognition of the NBA‐containing peptide presented in this study was successfully established. Such a combination of NBA and phage display technologies would be potential to efficiently confirm valuable target proteins binding specifically to capturing agents, to be arrayed onto solid surfaces to develop the designed peptide chip. © 2007 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 88: 131–140, 2007. This article was originally published online as an accepted preprint. The ‘Published Online’ date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com