Ribosomal Synthesis of Peptides with Unnatural Building Blocks

具有非天然结构单元的肽的核糖体合成

• 批准号: 8082393
• 负责人: PETER G SCHULTZ
• 金额: $ 35.59万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8082393
• 关键词: Affinity; Amino Acids; Bacteria; Biological; Chemicals; Complement; Cyclic Peptides; Development; Disease; Electronics; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Generations; Genetic; Libraries; Metalloproteases; Methodology; Methods; Molecular Structure; Mutagenesis; Mutation; Peptide Library; Peptide Synthesis; Peptides; Phage Display; Preparation; Property; Proteins; Serine Protease; System; Technology; Therapeutic; Therapeutic Uses; Time; base; functional group; in vivo; inhibitor/antagonist; intein; novel; novel strategies; protein function; research study

DESCRIPTION (provided by applicant): We will attempt to use our unnatural amino acid mutagenesis methodology to genetically encode libraries of cyclic and linear peptides containing a diverse array of unnatural amino acids. These peptide libraries will be generated both intracellularly in bacteria and in a phage-displayed format to allow in vivo selections and display-based affinity panning, respectively. This strategy will enable direct genetic encoding of molecular structure, random or directed mutagenesis, selection, and biological amplification, while at the same time allowing chemical diversity beyond what the canonical 20 amino acid repertoire can offer. The integration of unnatural amino acids into genetically encoded peptide libraries should facilitate both evolutionary experiments and the generation of peptides with novel biological activities that may ultimately find therapeutic use. This will be achieved through the following specific aims: 1. The preparation and incorporation of novel unnatural amino acids with unique reactivities and with functional groups capable of targeting specific proteins involved in disease states (e.g., serine proteases, metalloproteases, etc.) 2. The use of intein-based intracellular cyclic peptide synthesis methodologies together with unnatural amino acid technology to generate novel cyclic peptide libraries and select specific enzyme inhibitors 3. The use of phage display systems to generate and screen libraries of peptides containing unnatural amino acids for inhibitors of specific protein targets. PUBLIC HEALTH RELEVANCE: The ability to employ unnatural amino acids in biological selections and screens of peptide libraries should complement current synthetic and biological methods for generating biologically active peptides. Importantly, it will allow us to combine the power of templated biological synthesis, mutation, amplification, and selection with the chemist's ability to synthesize amino acids with defined steric and electronic properties not found in the canonical amino acids. The demonstration that these libraries can be efficiently selected or screened to identify specific inhibitors of protein function may ultimately provide a new approach to the development of peptide therapeutics.

描述（由申请人提供）：我们将尝试使用我们的非天然氨基酸诱变方法来遗传编码含有多种非天然氨基酸的环状和线性肽的文库。这些肽库将在细菌细胞内生成，并以噬菌体展示形式生成，以分别进行体内选择和基于展示的亲和淘选。这种策略将使分子结构的直接遗传编码、随机或定向诱变、选择和生物扩增成为可能，同时允许化学多样性超出典型的20个氨基酸库所能提供的范围。将非天然氨基酸整合到遗传编码的肽文库中应有助于进化实验和产生具有新生物活性的肽，这些肽最终可能会发现治疗用途。这将通过以下具体目标实现：1.制备和掺入具有独特反应性和具有能够靶向参与疾病状态的特定蛋白质（例如，丝氨酸蛋白酶、金属蛋白酶等）2.利用内含肽为基础的胞内环肽合成方法结合非天然氨基酸技术构建新型环肽文库并筛选特异性酶抑制剂3.使用噬菌体展示系统来产生和筛选含有非天然氨基酸的肽文库以用于特异性蛋白质靶点的抑制剂。 公共卫生相关性：在肽文库的生物选择和筛选中使用非天然氨基酸的能力应补充用于产生生物活性肽的当前合成和生物方法。重要的是，它将使我们能够将模板化生物合成、突变、扩增和选择的能力与化学家合成具有标准氨基酸所没有的空间和电子特性的氨基酸的能力联合收割机结合起来。这些文库可以被有效地选择或筛选以鉴定蛋白质功能的特异性抑制剂的证明可能最终为肽治疗剂的开发提供新的方法。