Role of anti-adhesin, podocalyxin and adaptor proteins in glomerular injury

抗粘附素、足萼蛋白和接头蛋白在肾小球损伤中的作用

• 批准号: 16590783
• 负责人: TAKEDA Tetsuro
• 金额: $ 2.37万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590783/
• 关键词: renal glomerulus; protein-protein interaction; phosphorylation; adaptor protein; podocyte

We have previously demonstrated that anti-adhesin molecule, podocalyxin binds to the second PDZ domain of NHERF2 and ezrin in vitro and in vivo and that the interaction is important to maintain the unique organization of podocyte structure. In this research project, we investigated how the adaptor proteins interact with podocalyxin in glomerular injury.1. We found that phosphrylation status of podocalyxin/NHERF2/ezrin comlex linked to the golmerular injury.2. We identified the protein which interacted with first PDZ domain of NHERF2 by yeast two-hybrid system. Surprisingly, it was NHERF2 itself. This self-interaction was confirmed by GST pull-down assay and co-immunoprecipitation and was disrupted in severe injured glomeruli.3. We analyzed the endocytic function of mouse immortalized glomerular epithlial cells that was established by Dr. Mundel like as in vivo millue.4. We also found that the interaction between podocalyxin and adaptor proteins of above cell line was reduced in propotion to the angioteinsin II concentration.5. When highly polarized MDCK cells were transfected to express podocalyxin, the adaptor proteins were re-localized to the apical submembranous domain and small G-protein, Rho A was activated. These results suggest podocalyxin may lead the adaptor proteins re-locate to the apical domain through the activation of Rho A.

我们以前已经证明，抗粘附素分子，足糖萼蛋白结合NHERF 2和ezrin在体外和体内的第二个PDZ结构域和相互作用是重要的，以维持足细胞结构的独特组织。在本研究项目中，我们研究了在肾小球损伤中衔接蛋白如何与足糖萼蛋白相互作用。我们发现足糖萼蛋白/NHERF 2/ezrin复合物的磷酸化状态与肾小球损伤有关.利用酵母双杂交系统鉴定了与NHERF 2第一个PDZ结构域相互作用的蛋白。令人惊讶的是，它是NHERF 2本身。GST pull-down和免疫共沉淀证实了这种自身相互作用，并且在严重损伤的肾小球中这种相互作用被破坏.我们分析了由Mundel博士建立的小鼠永生化肾小球上皮细胞的内吞功能，就像在体内一样。我们还发现足糖萼蛋白与上述细胞系衔接蛋白之间的相互作用随血管紧张素II浓度的增加而降低.当转染高度极化的MDCK细胞以表达足糖萼蛋白时，接头蛋白重新定位于顶端膜下结构域，并且小G蛋白Rho A被激活。这些结果表明podocalyxin可能通过激活Rho A而使接头蛋白重新定位到顶端结构域。

项目成果

「糸球体上皮細胞でのポドカリキシンの役割」Annual Review腎臓2004(伊藤克己、遠藤仁、御手洗哲也、東原英二、秋澤忠男編)

“足足萼蛋白在肾小球上皮细胞中的作用”2004 年肾脏年度评论（由 Katsumi Ito、Hitoshi Endo、Tetsuya Mitarai、Eiji Higashihara 和 Tadao Akizawa 编辑）

• 发表时间: 2004
• 作者: 竹田徹朗;下条文武
• 通讯作者: 下条文武

Significance of Proximal Tubular Metabolism of Advanced Glycation End Products in Kidney Diseases

• DOI: 10.1196/annals.1333.072
• 发表时间: 2005-06
• 期刊: Annals of the New York Academy of Sciences
• 影响因子: 5.2
• 作者: A. Saito;T. Takeda;Kiyoko Sato;Hitomi Hama;Atsuhito Tanuma;R. Kaseda;Yoshiki Suzuki;F. Gejyo

Podocalyxin activates RhoA and induces actin reorganization through NHERF1 and ezrin in MDCK cells

• DOI: 10.1097/01.asn.0000135968.49899.e8
• 发表时间: 2004-09-01
• 期刊: JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY
• 影响因子: 13.6
• 作者: Schmieder, S;Nagai, M;Farquhar, MG
• 通讯作者: Farquhar, MG

Evidence indicating that renal tubular metabolism of leptin is mediated by megalin but not by the leptin receptors

• DOI: 10.1210/en.2004-0074
• 发表时间: 2004-08-01
• 期刊: ENDOCRINOLOGY
• 影响因子: 4.8
• 作者: Hama, H;Saito, A;Gejyo, F
• 通讯作者: Gejyo, F

Evidence for megalin-mediated proximal tubular uptake of L-FABP, a carrier of potentially nephrotoxic molecules

• DOI: 10.1038/labinvest.3700240
• 发表时间: 2005-04-01
• 期刊: LABORATORY INVESTIGATION
• 影响因子: 5
• 作者: Oyama, Y;Takeda, T;Saito, A
• 通讯作者: Saito, A