Expressed Bacterial Triple-Helical Products as Tissue Engineering Scaffolds

表达细菌三螺旋产物作为组织工程支架

• 批准号: 7177983
• 负责人: BARBARA M BRODSKY
• 金额: $ 15.55万
• 依托单位: UNIV OF MED/DENT NJ-R W JOHNSON MED SCH
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-29 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7177983
• 关键词: basement membrane; binding sites; collagen; proteins; tissue engineering

DESCRIPTION (provided by applicant): The unique properties of the collagen triple-helix motif include its molecular hydrodynamic properties, extensive hydration, ability to bind diverse ligands, and the capacity self-associate to form fibrils and other higher order structures. These distinctive features have been exploited by nature to fill a wide range of structural and functional niches, and by scientists to design numerous products important for biomaterial and biomedical uses. Because of problems and concerns with the use of extracted collagens, attention has shifted to production of recombinant animal collagens, but efficient production has been slowed down addressing issues of post-translational modification. Rather than focus on specific types of animal collagens, we propose to focus on the intrinsic properties of the motif, using a bacterial triple-helix module which has high stability with no post-translational modifications. Stable triple-helix modules based on bacterial "collagen-like" sequences will be utilized to form triple-helical polymers in a high yield E. coli cold shock expression system. Repeating modules will be used to attain lengths comparable to that of animal fibril forming collagens, and we hypothesize that these triple-helical proteins will self-assemble to form fibrillar structures. Defined human collagen sequences involved in known interactions will be inserted between adjacent bacterial collagen modules to produce chimeras with functional binding sites. Introduction of basement membrane type breaks in the repeating (Gly-X-Y)n sequence between bacterial triple-helix modules will used to create a flexible or kinked type structure that more closely models basement membrane collagen networks and may provide a substrate for stem cell cultures. This novel approach plans to capture the flexibility and advantages of the collagen triple-helix motif while retaining the simplicity of a bacterial expression system. The products of this system will be ideal for manipulations and applications as scaffolds in tissue engineering. Lay Summary: A novel approach to gain efficient production of repeating bacterial collagen-like sequences plans to capture the flexibility and advantages of the collagen triple-helix motrf while retaining the simplicity of a bacterial expression system. The products of this system will be ideal for manipulations and applications as scaffolds in tissue engineering.

描述（由申请人提供）：胶原蛋白三螺旋基序的独特性质包括其分子流体动力学性质、广泛的水合作用、结合不同配体的能力以及自缔合形成原纤维和其他更高级结构的能力。这些独特的功能已被自然界利用，以填补广泛的结构和功能利基，并由科学家设计了许多重要的生物材料和生物医学用途的产品。由于使用提取的胶原蛋白的问题和担忧，注意力已经转移到重组动物胶原蛋白的生产上，但是由于翻译后修饰的问题，有效的生产已经减慢。而不是专注于特定类型的动物胶原蛋白，我们建议专注于基序的内在特性，使用细菌三螺旋模块，具有高稳定性，没有翻译后修饰。基于细菌“胶原蛋白样”序列的稳定三螺旋模块将用于以高产率E. coli冷休克表达系统。重复模块将被用来获得与动物原纤维形成胶原蛋白的长度相当的长度，我们假设这些三螺旋蛋白将自组装形成原纤维结构。参与已知相互作用的确定的人胶原序列将插入相邻的细菌胶原模块之间，以产生具有功能结合位点的嵌合体。在细菌三螺旋模块之间的重复（Gly-X-Y）n序列中引入基底膜型断裂将用于产生柔性或扭结型结构，其更紧密地模拟基底膜胶原网络，并且可以为干细胞培养物提供底物。这种新的方法计划捕获胶原蛋白三螺旋基序的灵活性和优势，同时保留细菌表达系统的简单性。该系统的产品将是理想的操作和应用的组织工程支架。简单总结：一种获得重复细菌胶原蛋白样序列的有效生产的新方法计划捕获胶原蛋白三螺旋motrf的灵活性和优点，同时保留细菌表达系统的简单性。该系统的产品将是理想的操作和应用的组织工程支架。