ENGINEERING PROTEINS WITH UNUSUAL ARCHITECTURES

工程蛋白质具有不寻常的结构

• 批准号: 2004274
• 负责人: JAMES P TAM
• 金额: $ 17.65万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1998-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2004274
• 关键词: chemical bond; cholecystokinin; cytokine receptors; defensins; drug design /synthesis /production; guinea pigs; helper T lymphocyte; immunomodulators; inhibitor /antagonist; laboratory mouse; malaria vaccines; neuropeptides; peptide chemical synthesis; protein engineering; protein structure function; synthetic vaccines; vaccine development

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.

该应用程序将重点关注方法的开发和 具有不寻常结构的蛋白质的治疗应用。已选择 我们设计的蛋白质将被环化和分支蛋白质。目标 环化蛋白包括白细胞介素-1受体拮抗剂，它是 目前正在临床试验中作为一种药物来减轻脓毒症的严重程度 关节炎;监测肽，它是一种胆囊收缩素释放因子， 可能有助于治疗消化系统疾病；和防御素是 一种广谱抗生素，具有对抗艾滋病相关活性的良好前景 病原体。目标分支蛋白将包括疟疾疫苗 含有源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。这 分支设计将 MSP-1 抗原的多聚体副本纳入 小肽基核心基质作为多抗原肽（MAP）。这 MSP-1 衍生的抗原具有构象依赖性，包含多个 二硫键。与 T 辅助表位和内置佐剂一起 组装成 MAP，它将提供一种完整的疫苗 明确的结构。这种疟疾疫苗将接受保护测试 Carol Long 博士在小鼠模型中对抗感染挑战 疟疾领域的顶尖专家。含有 HIV-1 抗原的 MAP 版本是 目前作为候选艾滋病疫苗正在进行一期临床试验。 该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域连接策略。 它是一种连接或环化完全未受保护的肽和 通过肽键形成蛋白质片段，无需激活。这个方法是 非常适合合成环化和分支蛋白质， 无法直接通过重组 DNA 方法获得，并且 传统的肽合成方法很难获得。这 结构域连接策略是有机和肽的结合方法 用于治疗应用的工程蛋白质化学。