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

Cell surface engineering

细胞表面工程

基本信息

批准号：
10145107
负责人：
TANAKA Atsuo
金额：
$ 57.09万
依托单位：
Kyoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas (A)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2001
项目状态：
已结题

项目摘要

In cell surface, many recepter proteins are presented and the specific biomolecules are recognized by these recepter proteins. These mechanisms can be analyzed from chemical view points. To systemize the biotergeting, the cell sueface design was investigated. Dr. Tanaka studied the development of novel yeast cells armed with enzymes and functional proteins (glucoamylase, α-amylase, CM-cellulase, β-glucosidase, lipase, a histidine oligomer. green fluorescent protein, and its variants). The yeasts were constructed by a cell surface engiseering system of yeast Sacebaromyces cerevisiae, which can assimilate the redundant biomass or can display the vaccine protein. These surface-engineered yeast cells are termed as "Arming yeasts". "Cell surface engineering" will be capable of conferring novel additional abilities upon living cells and will herald a new era in the field of biotechnology. Dr. Nagamune Studied a way of changing the binding element of a growth factor so as to alter its specificity of specific ligand-receptor pairs. By joining these antibody variable domains, VH and VL, of anti-hen egg lysozyme (HEL) antibody HyHEL-10 to the cytoplasmic portion of erythropoietin receptor (EpoR), a chimeric receptor that could he activated by HEL was developed. When IL-3-dependent murine myeloid 32D cells were co-transfected with plasmids encoding VH-EpoR and VL-EpoR-IRES-EGFP, HEL induced selective expansion of the cell population with high expression level of EGFP. To expand the current approach, chimeric receptors whose cytoplasmic domain was replaced by that of gp 130 were developed. Dr. Akiyoshi studied the functions of cell-surface saccharides using liposomal displaying systems as model biological membrane. It was demonstrated that 1) function of gangliosides-conatining liposomes, 2) coating of phytylphosphate giant vesicles with a hydrophobized polysaccharide 3) dynamic morphological changes of cell-size liposomes in the presence of gangliosides.

在细胞表面，存在许多受体蛋白，这些受体蛋白识别特定的生物分子。这些机制可以从化学角度进行分析。为了使生物接合系统化，研究了细胞表面设计。Tanaka博士研究了新型酵母细胞的开发，这些酵母细胞配备了酶和功能性蛋白质（葡糖淀粉酶、α-淀粉酶、CM-纤维素酶、β-葡萄糖苷酶、脂肪酶、组氨酸寡聚物）。绿色荧光蛋白及其变体）。利用酿酒酵母的细胞表面工程系统构建了能消化多余生物量或展示疫苗蛋白的酵母。这些表面工程化的酵母细胞被称为“武装酵母”。“细胞表面工程”将能够赋予活细胞新的额外能力，并将预示着生物技术领域的一个新时代。Nagamune博士研究了一种改变生长因子的结合元件的方法，以改变其特异性配体-受体对的特异性。通过将抗鸡蛋溶菌酶（HEL）抗体HyHEL-10的这些抗体可变结构域VH和VL连接到促红细胞生成素受体（EpoR）的胞质部分，开发了可被HEL激活的嵌合受体。将编码VH-EpoR和VL-EpoR-IRES-EGFP的质粒共转染IL-3依赖的小鼠髓系32 D细胞，HEL诱导具有高水平EGFP表达的细胞群体的选择性扩增。为了扩展目前的方法，嵌合受体的胞质结构域被替换的糖蛋白130的开发。Akiyoshi博士使用脂质体展示系统作为模型生物膜研究细胞表面粘附的功能。结果表明：（1）含有神经节苷脂的脂质体的功能;（2）用疏水化多糖包被植脂酰磷酸巨囊泡;（3）神经节苷脂存在下细胞大小的脂质体的动态形态变化。