Usui Kenji, Tomizaki Kin-Ya, Ohyama Takafumi, Nokihara Kiyoshi, Mihara Hisakazu
Department of Bioengineering and the COE21 program, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Nagatsuta-cho 4259 B-40, Midori-ku, Yokohama 226-8501, Japan.
Mol Biosyst. 2006 Feb;2(2):113-21. doi: 10.1039/b514263f. Epub 2005 Dec 23.
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.
一种新型的干肽微阵列系统已被构建出来，它为蛋白质检测和分析提供了一种切实可行的解决方案。该系统是一种在干燥条件下的阵列制备和检测程序，使用设计好的肽作为非固定化捕获剂来检测蛋白质。该系统具有几个优点，包括其便携性和易用性，以及无需考虑样品溶液蒸发的问题。在本研究中，利用一个α-螺旋肽微型文库对各种蛋白质进行了表征。当将蛋白质添加到肽文库阵列中时，荧光肽根据其序列显示出不同的荧光强度。这些响应模式可被视为“蛋白质指纹”（PFPs），足以确定目标蛋白质的身份。此外，使用聚类分析对所得的PFPs进行了统计分析。根据蛋白质的家族和结合特性，成功地对蛋白质的PFPs进行了聚类。此外，使用纳升系统对目标蛋白质进行了表征，检测下限可达1.2飞摩尔。这些研究表明，干肽阵列系统是一种用于检测和分析目标蛋白质的有前途的工具。干肽阵列将在用于蛋白质组学和配体筛选研究的高通量蛋白质检测纳米/微阵列的开发中发挥作用。
