Design, expression, and properties of rHbs for HBOCs

HBOC 的 rHb 的设计、表达和特性

• 批准号: 6654243
• 负责人: CHIEN HO
• 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/6654243
• 关键词: Bohr effect; Escherichia coli; biomaterial development /preparation; biotechnology; blood /plasma substitute; dextrans; fluidity; hemoglobin; nitric oxide; nitroso compounds; nuclear magnetic resonance spectroscopy; oxygen transport; peptide chemical synthesis; polyethylene glycols; protein engineering; protein structure function; proteomics; recombinant proteins

DESCRIPTION (provided by applicant): The goal of this project is to design and express novel human mutant recombinant hemoglobins (rHbs), which, after reacting with polyethylene glycol (PEG) or other cross-linking reagents, possess appropriate structural and functional properties to serve as hemoglobin-based oxygen carries (HBOCs). The vasoactivity associated with the use of many of the current HBOCs has challenged previous beliefs as to the optimal O2 affinity, cooperatively, Bohr effect, size, viscosity, and other properties needed for HBOCs. rHbs that we have previously prepared and new ones that we will design and express will be used to test both the new paradigms for the design of blood substitutes and the various hypotheses regarding the origin of vasoactivity. Using our Hb expression system in Escherichia coli, we are in a position to design and express any rHbs needed. Our laboratory has extensive experience and is also fully equipped to carry out structural studies using nuclear magnetic resonance (NMR) and functional studies using biochemical-biophysical techniques (e.g., equilibrium oxygen binding, kinetics of ligand binding, etc)on Hbs. A unique feature of this Program Project is the interactions among its component parts. The key investigators of this Program Project have worked together and have published papers together during the past ten years. The scope of this Program Project requires a multidisciplinary approach to the design of novel HBOCs. The structural and biochemical-biophysical results obtained from this project (Project 2) will provide new insights for Projects 1, 3, 4 and 5 as well as the Protein Biochemistry Core. Equally, the results from other components of the Program Project will guide Project 2 in designing the rHbs most effective as HBOCs. We believe that the results obtained from this Program Project will provide new knowledge for the development of safe, reliable HBOCs as blood substitutes.

描述（由申请人提供）： 本项目的目标是设计和表达新型的人突变重组血红蛋白 （rHbs），其在与聚乙二醇（PEG）或其它交联剂反应后， 具有适当的结构和功能特性，可用作血红蛋白基氧 携带（HBOCs）。与使用许多当前的HBOC相关的血管活性已经 挑战了以前的信念，如最佳的O2亲和力，合作，玻尔效应，大小， 粘度和HBOCs所需的其他性能。我们之前准备的rHbs， 我们将设计和表达的新模型将用于测试 血液替代品的设计和关于血管活性起源的各种假设。 使用我们的Hb表达系统在大肠杆菌中，我们能够设计和表达 需要的rhb我们的实验室拥有丰富的经验，也是设备齐全， 使用核磁共振（NMR）进行结构研究， 生物化学-生物物理技术（例如，平衡氧结合，配体动力学 结合等）。这个项目的一个独特之处是它的相互作用。 零部件该项目的主要研究人员共同努力， 在过去的十年里一起发表论文。本计划项目的范围 需要多学科的方法来设计新颖的HBOC。的结构和 从该项目（项目2）获得的生物化学-生物物理结果将为项目1，3，4和5以及蛋白质生物化学核心提供新的见解。同样，其他结果 计划项目的组成部分将指导项目2设计最有效的rHbs， HBOCs。我们相信，从这个项目中获得的结果将提供新的 开发安全、可靠的HBOCs作为血液替代品的知识。