权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

The development of next-generation albumin preparation by mutagenesi s studies

诱变研究开发下一代白蛋白制剂

基本信息

批准号：
14370759
负责人：
OTAGIRI Masaki
金额：
$ 8.58万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

Human serum albumin(HSA) has many essential functions for homeostasis, such as the maintenance of osmotic pressure, drug binding and antioxidant activity. Mutagenesis studies of HSA made it possible to examine the participation of various amino acid residues and domains in the functional properties of HSA, such as binding capacity, antioxidant activity and prolonged half-life.In this study, firstly, we studied the antioxidant properties of single-residue mutants of human serum albumin. Cysteine residues, especially ^<166>Cys contribute much to this function of albumin. Secondly, Pichia pastoris has been used to express each of the three rHSA domains separately. Recombinant domain I protein exhibited antioxidant activity comparable to that of rHSA. A novel domain exchange HSA with potentially high antioxidant activity was designed where three domain I proteins of HSA were genetically fused. This exchanged HSA had higher antioxidant activity than wild-type HSA. Thirdly, rHSA dimer has been expressed using Pichia pastoris. The structure and binding capacity of rHSA dimer was almost identical to that of native HSA. rHSA dimer was shown to exhibit better intravascular retention and lower vascular permeability properties. Lastly, single-residue mutants R410C efficiently introduced NO further than the wild type. S-NO-R410C had higher antibacterial activity than S-NO-HSA.The results of this research serve as important fundamental data for further development of new recombinant albumin.

人血清白蛋白（HSA）具有许多维持体内平衡的重要功能，如维持渗透压、药物结合和抗氧化活性。人血清白蛋白（HSA）的突变研究使人们有可能研究各种氨基酸残基和结构域参与HSA的功能特性，如结合能力、抗氧化活性和延长半衰期。半胱氨酸残基，特别是α<166>-Cys对白蛋白的这种功能有很大贡献。其次，巴斯德毕赤酵母已被用于分别表达三个rHSA结构域中的每一个。重组结构域I蛋白具有与rHSA相当的抗氧化活性。设计了一种具有潜在高抗氧化活性的新型结构域交换HSA，其中HSA的三个结构域I蛋白被基因融合。这种交换的HSA比野生型HSA具有更高的抗氧化活性。第三，已使用毕赤酵母表达rHSA二聚体。rHSA二聚体的结构和结合能力与天然HSA几乎相同。rHSA二聚体显示出更好的血管内滞留和更低的血管渗透性。最后，单残基突变体R410 C有效地引入NO进一步比野生型。S-NO-R410 C的抗菌活性高于S-NO-HSA，为进一步开发新型重组白蛋白提供了重要的基础数据。