ENGINEERING PROTEINS WITH UNUSUAL ARCHITECTURES

该应用程序将侧重于方法的发展， 具有不寻常结构的蛋白质的治疗应用。选择 用于我们设计的蛋白质将是环化和分支蛋白质。目标 环化蛋白包括白细胞介素-L受体拮抗剂， 目前正在临床试验中作为一种药物，以减少败血症的严重程度， 关节炎;监测肽是胆囊收缩素释放因子， 可用于治疗消化系统疾病;和防御素， 一种广谱抗生素，对艾滋病相关的 病原体目标分支蛋白质将包括疟疾疫苗 含有来自裂殖子表面蛋白的保护性抗原， （MSP-1）。该抗原是迄今为止最有希望的候选疫苗。的 分支设计将MSP-1抗原的多聚体拷贝掺入到多聚体中。 小肽基核心基质作为多抗原肽（MAP）。的 来源于MSP-1的抗原是构象依赖性的， 二硫键与辅助性T细胞表位和内置佐剂一起 组装形成地图，它将提供一个完整的疫苗， 明确的结构。这种疟疾疫苗将进行保护试验 Carol Long博士在小鼠模型中对抗感染挑战， 疟疾方面的顶尖专家含有HIV-1抗原的MAP的一种形式是 目前作为候选艾滋病疫苗正在进行I期临床试验。 本应用程序的统一主题是开发和应用 我们实验室最近概念化的结构域连接策略。 它是一种连接或环化完全未保护的肽的方法， 蛋白质片段通过肽键而不活化。该方法 非常适合于合成环化和分支蛋白， 不能直接通过重组DNA方法获得， 难以通过常规肽合成方法获得。的 结构域连接策略是有机和肽的组合方法 化学工程蛋白质用于治疗应用。

This application will focus on the methodological development and therapeutic applications of proteins with unusual architectures. Selected proteins for our design will be circularized and branch proteins. Target circularized proteins include interleukin-l receptor antagonist which is currently in clinical trials as a drug to reduce severity of sepsis and arthritis; monitor peptide which is a cholecystokinin-releasing factor and may be useful for treatment of digestive disorders; and defensin which is a broad-spectrum antibiotic with promising activity against AIDS-related pathogens. Target branch proteins will include a malaria vaccine containing the protective antigen derived from merozoite surface protein (MSP-1). This antigen is the most promising vaccine candidate to date. The branch design will incorporate multimeric copies of MSP-1 antigen in a small peptidyl core matrix as multiple antigen peptides (MAPs). The antigen derived from MSP-1 is conformation-dependent and contains multiple disulfide bonds. Together with a T-helper epitope and a built-in adjuvant assembled to form MAPs, it will provide a complete vaccine with an unambiguous structure. This malaria vaccine will be tested for protection against infection challenge in the mouse model by Dr. Carol Long who is a leading expert in malaria. A version of MAPs containing a HIV-1 antigen is currently in Phase I clinical trials as a candidate AIDS vaccine. The unifying theme of this application is the development and application of the domain ligation strategy recently conceptualized in our laboratory. It is a method to link or circularize totally unprotected peptide and protein segments via a peptide bond without activation. This method is well suited for the synthesis of circularized and branch proteins which are inaccessible directly by the recombination DNA method and are difficult to obtain by the conventional peptide synthesis approaches. The domain ligation strategy is a combined approach of organic and peptide chemistry in engineering proteins for therapeutic applications.