Kubota Satoshi, Kawaki Harumi, Takigawa Masaharu
Advanced Research Center for Oral and Craniofacial Sciences, Okayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, 2-5-1 Shikata-cho, Kita-ku, Okayama, 700-8525, Japan.
Department of Membrane Biochemistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama, Japan.
Methods Mol Biol. 2017;1489:155-167. doi: 10.1007/978-1-4939-6430-7_16.
Function of CCN family proteins is determined by the interactions with multiple cofactors that are present in microenvironment. Therefore, finding out these cofactors is critically important in understanding the molecular function of the CCN family members. For this objective, bacteriophage random peptide display library is a quite feasible tool. In this library, each filamentous bacteriophage is designed to display an oligopeptide of random 12-16 amino acid residues on its surface. Bacteriophage clones that possess the peptides that bind to a CCN family protein are selected through several cycles of a process designated biopanning or affinity selection. By determining the nucleotide sequence of the DNA that encodes the displayed peptide, oligopeptides that specifically bind to the CCN family member can be specified. Obtained peptide sequences can be utilized for designing peptide aptamers for the CCN family protein, or as a key sequence to find out new CCN family cofactor candidates in silico.
CCN家族蛋白的功能由其与微环境中多种辅因子的相互作用所决定。因此，找出这些辅因子对于理解CCN家族成员的分子功能至关重要。为此，噬菌体随机肽展示文库是一个非常可行的工具。在这个文库中，每个丝状噬菌体被设计成在其表面展示一个由12 - 16个氨基酸残基组成的随机寡肽。通过称为生物淘选或亲和选择的几个循环过程，筛选出携带与CCN家族蛋白结合肽的噬菌体克隆。通过确定编码所展示肽的DNA的核苷酸序列，可以明确与CCN家族成员特异性结合的寡肽。获得的肽序列可用于设计CCN家族蛋白的肽适配体，或作为在计算机上寻找新的CCN家族辅因子候选物的关键序列。
