Core--Protein biochemistry

核心--蛋白质生化

• 批准号: 6654250
• 负责人: BELUR N MANJULA
• 金额: $ 38.74万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-05 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6654250
• 关键词: analytical chemistry; biochemistry; biomaterial development /preparation; biomedical facility; biotechnology; blood /plasma substitute; hemoglobin; peptide chemical synthesis; protein purification; protein structure function; sample collection; tissue /cell preparation

DESCRIPTION (provided by applicant): The Protein Biochemistry Core performs the important function of integrating the design strategies of Projects 1 and 2 to produce a blueprint of a matrix of products that can be used for validating the new design paradigms proposed in the Program Project. The success of the individual projects of the Program Project depends heavily on the availability of the various site-specifically surface decorated and inter-tetramerically crosslinked Hbs. This is the responsibility of the Protein Biochemistry Core. The major Core products are 1) Tetrameric Hbs - unmodified HbA mutant Hbs-chemically modified high and low oxygen affinity Hbs, and 2) size-enhancedsHbS namely, the surface decorated Hbs-PEGylated and dextranylated Hbs, and oligomerized Hbs. The native or mutagenized or chemically modified tetrameric Hbs will serve as the starting materials for preparation of the size enhanced Hbs. The Protein Biochemistry Core will receive information from Project 1 on the methodologies for the preparation of the various PEGylated, dextranylated and oligomeric Hbs. A selected set of protocols will be applied to the recombinant Hbs developed in Project 2. Initially, the protocols will be scaled up to generate materials at 1 - 2 g levels. The products generated will be purified and subjected to a variety of biochemical characterization analyses. The materials generated will also be analyzed for their oxygen binding characteristics and supplied to Dr. Vandegriff (Sangart) for characterization of their physical properties- viscosity and colloidal osmotic pressure, and to Projects 2, 3 and 4 for Biophysical investigations. As an initial screen, the materials exhibiting viscosity, COP and hydrodynamic volume comparable to P5K6-HbA will be supplied to Project 5 for screening of their vasoactivity by the top load assay in hamster. The integration of the results of these studies with those from Projects 2, 3 and 4 will lead to the identification of a set of materials for more detailed investigations in the microcirculatory assay (exchange transfusion and hemorrhagic shock models), and for measurements of diffusive oxygen transport in artificial capillaries. Such materials will be generated at 5 - 10 g levels. The Protein Biochemistry Core will coordinate with the BioAffinity Systems for the large-scale synthesis of the PEG reagents. The Core will also provide support for Projects 3 and 4 in the synthesis of specialty -probe labeled surface decorated Hbs for studies on the local and global effects of the surface decoration on the structure of Hb.

描述（由申请人提供）： 蛋白质生物化学核心执行集成设计的重要功能 项目1和2的策略生产可以使用的产品矩阵的蓝图蓝图 用于验证程序项目中提出的新设计范式。成功的成功 该计划项目的各个项目在很大程度上取决于各种位点特定地面装饰和四个交联的HBS的可用性。这是蛋白质生物化学核心的责任。主要的核心产品是1）四聚体HBS-未修饰的HBA突变体HBS化学改性的高和低氧亲和力HBS和2）尺寸增强的shangbass，即表面装饰的HBS-pegegypated和右旋烷基化的HBS，以及 低聚的HBS。天然或诱变或化学修饰的四聚体HBS将使用 作为准备大小增强的HB的起始材料。蛋白质生物化学 核心将从项目1中接收有关准备方法的信息 各种高乙二醇，右烷基化和低聚的HBS。选定的一组协议将是 应用于项目2中开发的重组HBS。最初，协议将缩放 以1-2 g的水平生成材料。生成的产品将被纯化并进行 各种生化特征分析。生成的材料也将是 分析其氧结合特征，并提供给Vandegriff博士（Sangart） 其物理特性的表征 - 粘度和胶体渗透压，以及 项目2、3和4进行生物物理研究。作为初始屏幕，展示的材料 将提供与P5K6-HBA相当的粘度，COP和流体动力量 5用于通过仓鼠中的最高负荷测定法筛选其血管活性。整合 这些研究与项目2、3和4的研究的结果将导致鉴定一组材料，以在微循环测定中进行更详细的研究（交换输血 和出血性休克模型），用于测量人造中的扩散氧运输 毛细血管。此类材料将在5-10 g水平下产生。蛋白质生物化学核心将 与生物障碍系统协调，以大规模合成PEG试剂。这 核心还将为项目3和4的项目提供支持。 表面装饰的HB用于研究表面装饰对局部和全球影响的研究 HB的结构。