Characterization of β cell-specific gene obtained by large cDNA sequencing.

通过大型 cDNA 测序获得的 β 细胞特异性基因的表征。

• 批准号: 15590943
• 负责人: YOSHIMOTO Katsuhiko
• 金额: $ 2.24万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15590943/
• 关键词: pancreatic islet β cells; serine protease; endosome

To characterize expressed sequence tag (EST) profiling in pancreatic islet β cells, a large-scale cDNA sequencing analysis was carried out for ESTs derived from MIN6 cells, a cell line of murine pancreatic islet β cells. Among these, 3 individual ESTs were found to code one novel protein which we termed isletasin. Isletasin gene encoded 552 amino acids. Isletasin contained repeated catalytic triads for S1 family of serine proteases, and its expression was limited to MIN6 cells or mouse pancreatic islets. Rat homologue was cloned from INS-1 cell line. Identity of amino acids between mouse and rat was 94.8%. Real-time RT-PCR showed high expression of isletasin in pancreatic β cell lines of MIN6, βHC9, INS-1 and pancreatic islets isolated from mice and rats. Immunoblotting of extract from COS-7 cells expressing epitope-tagged isletasin showed that isletasin is not an integral but a peripheral protein. Isletasin is strongly associated with the luminal surface of the microsomal membrane, where it undergoes signal peptide cleavage and N-glycosylation. The fluorescence detection of each organelle marker protein and green fluorescence protein-tagged isletasin or His, myc-tagged isletasin in MIN6 cells revealed that isletasin is an early endosomal protein.

为了表征胰岛β细胞中的表达序列标签（EST）谱，对来自小鼠胰岛β细胞的细胞系MIN 6细胞的EST进行大规模cDNA测序分析。其中，3个独立的EST被发现编码一种新的蛋白质，我们称之为isletasin。Isletasin基因编码552个氨基酸。Isletasin含有丝氨酸蛋白酶S1家族的重复催化三联体，其表达仅限于MIN 6细胞或小鼠胰岛。从INS-1细胞系克隆大鼠同源物。小鼠与大鼠氨基酸同源性为94.8%。Real-time RT-PCR结果显示胰岛β细胞株MIN 6、β HC 9、INS-1及小鼠、大鼠胰岛均表达Isletasin。来自表达表位标记的isletasin的COS-7细胞的提取物的免疫印迹显示isletasin不是一个整体而是一个外周蛋白。Isletasin与微粒体膜的腔表面密切相关，在那里它经历信号肽切割和N-糖基化。在MIN 6细胞中对每个细胞器标记蛋白和绿色荧光蛋白标记的isletasin或His，myc标记的isletasin的荧光检测显示isletasin是早期内体蛋白。

项目成果

Retinoic acid disintegrated desmosomes and hemidesmosomes in stratified oral keratinocytes

• DOI: 10.1111/j.1600-0714.2004.00245.x
• 发表时间: 2004-11-01
• 期刊: JOURNAL OF ORAL PATHOLOGY & MEDICINE
• 影响因子: 3.3
• 作者: Hatakeyama, S;Hayashi, S;Satoh, M
• 通讯作者: Satoh, M

Redox State-Dependent and Sorbitol Accumulation-Independent Diabetic Albuminuria in Mice with Transgene-Derived Human Aldose Reductase and Sorbitol Dehydrogenase Deficiency

转基因衍生的人醛糖还原酶和山梨醇脱氢酶缺陷小鼠中氧化还原状态依赖性和山梨醇积累非依赖性糖尿病蛋白尿

• 发表时间: 2004
• 期刊: Diabetologia 47
• 作者: Ii S;et al.
• 通讯作者: et al.

Differentiation phenotypes of pancreatic islet β- and α-cells are closely related with homeotic genes and a group of differentially expressed genes

• DOI: 10.1016/j.gene.2004.01.016
• 发表时间: 2004-04-28
• 期刊: GENE
• 影响因子: 3.5
• 作者: Mizusawa, N;Hasegawa, T;Yoshimoto, K
• 通讯作者: Yoshimoto, K

Redox State-Dependent and Sorbitol Accumulation-Independent Diabetic Albuminuria in Mice with Transgene-Derived Human Aldose Reductase and Sorbitol Dehydrogenase Deficiency.

转基因衍生的人醛糖还原酶和山梨醇脱氢酶缺乏的小鼠中氧化还原状态依赖性和山梨醇积累非依赖性糖尿病蛋白尿。

• 发表时间: 2004
• 期刊: Diabetologia 47(3)
• 作者: Ii S;et al.
• 通讯作者: et al.

Gamma-tubulin in Basal Land Plants : Characterization, Localization, and Impliction in the Evolution of Acentriolar Microtubule Organizing Centers.

基底陆地植物中的γ-微管蛋白：心轴微管组织中心进化的特征、定位和含义。

• 发表时间: 2004
• 期刊: Plant Cell 16
• 作者: Hatakeyama S;et al.;Srtsuko Ii et al.;Noriko Mizusawa et al.;Shimamura M et al.
• 通讯作者: Shimamura M et al.