Design, expression, and properties of rHbs for HBOCs
Design, expression, and properties of rHbs for HBOCs
批准号:
6654243
负责人:
CHIEN HO
金额:
$38.74万
依托单位国家:
美国
项目类别:
财政年份:
2002
资助国家:
美国
项目状态:
已结题
起止时间:
2002-08-05 至 2007-07-31
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
中文摘要
描述(由申请人提供):
本项目的目标是设计和表达新型人类突变型重组血红蛋白。
(RHbs),其在与聚乙二醇(PEG)或其他交联剂反应后,
具有适当的结构和功能性质,可用作基于血红蛋白的氧
携带(HBOC)。与当前许多HBOC的使用相关的血管活性
挑战了之前关于最佳O2亲和力的信念,协同作用,玻尔效应,大小,
粘度,以及HBOC所需的其他性质。我们之前准备的rHb和
我们将设计和表达的新范例将用于测试
血液替代品的设计和关于血管活性起源的各种假说。
利用我们的Hb在大肠杆菌中的表达系统,我们能够设计和表达
任何需要的rHb。我们的实验室经验丰富,设备齐全,可以进行
我们使用核磁共振进行结构研究,并使用功能研究
生化-生物物理技术(例如,平衡氧结合、配体的动力学
绑定等)。该计划项目的一个独特特点是其
部件。该计划项目的主要研究人员已经合作并
在过去的十年里一起发表了论文。本计划项目的范围
需要一个多学科的方法来设计新颖的HBOC。结构和
从该项目(项目2)获得的生化-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,来自其他方面的结果
计划项目的组成部分将指导项目2设计最有效的rHBS AS
HBOC。我们相信,从该计划项目中获得的结果将提供新的
开发安全、可靠的HBOC作为血液替代品的知识。
英文摘要
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.
期刊论文(0)
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