Protocols for screening peptides binding to CCN family proteins and their extended utility
Protocols for screening peptides binding to CCN family proteins and their extended utility
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筛选与 CCN 家族蛋白结合的肽的方案及其扩展用途
DOI：
10.1007/978-1-0716-2744-0_8
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Takigawa M
中科院分区：
文献类型：
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作者：
Kubota S;Kawaki H;Takigawa M
CCN 家族蛋白的功能取决于它们与微环境中存在的多种辅因子的相互作用。因此，确定这些辅助因子对于理解 CCN 家族成员的分子功能至关重要。对于这个目标，噬菌体随机肽展示库是一个合适的工具。在这个文库中，每个丝状噬菌体都被设计为在其表面展示由 7-20 个随机氨基酸残基组成的寡肽。具有与 CCN 家族蛋白结合的肽的噬菌体克隆是通过称为生物淘选或亲和选择的过程的几个循环来选择的。通过确定编码展示的肽的DNA的核苷酸序列，可以指定特异性结合CCN家族成员的寡肽。获得的肽序列可用于设计CCN家族蛋白的肽适体，或作为关键序列在计算机上确定新的CCN家族辅因子候选物。可以使用来自幼稚淋巴细胞或来自 CCN 家族蛋白免疫的 B 细胞的抗体 cDNA 文库来代替随机肽 cDNA 文库，以获得高度特异性的 CCN 家族检测或功能调节工具。
The function of CCN family proteins is determined by their interactions with multiple cofactors that are present in the microenvironment. Therefore, determining these cofactors is critically important in understanding the molecular function of CCN family members. For this objective, a bacteriophage random peptide display library is a suitable tool. In this library, each filamentous bacteriophage is designed to display an oligopeptide of 7–20 random amino acid residues on its surface. Bacteriophage clones that possess peptides that bind to a CCN family protein are selected through several cycles of a process called biopanning or affinity selection. By determining the nucleotide sequence of the DNA that encodes the displayed peptide, the oligopeptides that specifically bind to the CCN family member can be specified. The obtained peptide sequences can be utilized to design peptide aptamers for CCN family proteins, or as a key sequence to determine new CCN family cofactor candidates in silico. Instead of a random peptide cDNA library, an antibody cDNA library from naïve lymphocytes or from B cells immunized by a CCN family protein can be used in order to obtain a highly specific CCN family detection or functional modulation tool.