Analysis of the physiological function of a newly discovered protein, parchorin, specifically expressed in water-secreting cells

分析新发现的蛋白质，parchorin，在泌水细胞中特异性表达的生理功能

• 批准号: 13470511
• 负责人: URUSHIDANI Tetsuro
• 金额: $ 8万
• 依托单位: The University of Tokyo.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470511/
• 关键词: parchorin; water movement; CLIC family protein; MDCK cells; casein kinase II; duct; purinergic; リン酸化; カゼインキナーゼ; Xenopus Oocytes

This work was conducted to elucidate the physiological function of parchorin, which we recently purified and cloned. Parchorin is specifically expressed in cells participating water movement in various tissues, I.e., ductal cells of exocrine glands, ciliary body, inner ear, renal tubules, etc. The amount of parchorin in the gastric mucosa, but not in the submandibular glands, increased after weaning. In the mammary gland, parchorin expression was greater in a lactating rabbit compared with a pregnant rabbit. The cellular distribution and changes in expression indicate that parchorin plays an important role in the cells participating water movement. Parchorin was suggested to translocate from cytosol to plasma membrane in association with stimulation. To elucidate the translocation mechanism, we developed a model system where GFP-parchorin was expressed in MDCK cells. When confluent cells were stimulated with ATP or bradykinin, parchorin translocated to the plasma membrane by calcium and protein kinase C-mediated pathway. We also found casein kinase II phosphorylates parchorin on its Ser83, 338, and 393. Mutational analysis of parchorin revealed that the phosphorylation of these residues resulted in the stabilization of the protein.

这项工作的目的是阐明我们最近纯化和克隆的 parchorin 的生理功能。 Parchorin特异表达于各种组织中参与水运动的细胞，即外分泌腺、睫状体、内耳、肾小管等的导管细胞。断奶后，胃粘膜中的Parchorin含量增加，但下颌下腺中的Parchorin含量不增加。在乳腺中，与怀孕的兔子相比，哺乳期的兔子的乳腺蛋白表达更高。细胞分布和表达变化表明parchorin在细胞参与水分运动中发挥着重要作用。据认为，Parchorin 从细胞质转移到质膜与刺激有关。为了阐明易位机制，我们开发了一个模型系统，其中 GFP-parchorin 在 MDCK 细胞中表达。当汇合细胞受到 ATP 或缓激肽刺激时，parchorin 通过钙和蛋白激酶 C 介导的途径转移到质膜。我们还发现酪蛋白激酶 II 在 Ser83、338 和 393 上磷酸化 parchorin。parchorin 的突变分析表明这些残基的磷酸化导致了蛋白质的稳定。

项目成果

J.Matsukawa, K.Tashiro, T.Nagao, T.Urushidani: "Role of small GTP-binding proteins and cytoskeleton in gastric acid secretion"Inflammopharmacol.. (in press). (2003)

J.Matsukawa、K.Tashiro、T.Nagao、T.Urushidani：“小 GTP 结合蛋白和细胞骨架在胃酸分泌中的作用”Inflammopharmacol..（出版中）。

Y. Mizukawa, T. Nishizawa, T. Nagao, K. Kitamura, and T. Urushidani , ed. by T. Urushidani, J. G. Forte,and G. Sachs: "Tissue and Cell Distribution of Parchorin, a Chloride Intracellular Channel-Related Protein in Mechanisms And Consequences of Proton Tra

Y. Mizukawa、T. Nishizawa、T. Nagao、K. Kitamura 和 T. Urushidani 编辑。

Shanks RA, Larocca MC, Benyman M, Edwards JC, Urushidani T, Navarre J, Goldenring JR: "AKAP350 at the Golgi apparatus. II. Association of AKAP350 with a novel chloride intracellular channel (CLIC) family member"J. Biol. Chem.. 277. 40973-40980 (2002)

Shanks RA、Larocca MC、Benyman M、Edwards JC、Urushidani T、Navarre J、Goldenring JR：“高尔基体上的 AKAP350。II. AKAP350 与新型氯离子细胞内通道 (CLIC) 家族成员的关联”J.

T. Urushidani , ed. by T. Urushidani, J. G. Forte, and G. Sachs: "Permeabilized Gastric Gland Models ― A New Strategy for Analyzing Parietal Cell Signal Transduction" in "Mechanisms And Consequences of Proton Transport"Kluwer Academic. 209-220 (2002)

T. Urushidani，编辑：T. Urushidani、J. G. Forte 和 G. Sachs：“透化胃腺模型——分析壁细胞信号转导的新策略”，《质子传输的机制和后果》，Kluwer 学术期刊 209-220。 (2002)

N.Omi, T.Nagao, T.Urushidani: "Phosphatidylinositol is essential determinant for K+ permeability involved in gastric proton pumping"Am. J. Physiol. Gastrointest. Liver Physiol.. 281. G786-G797 (2001)

N.Omi、T.Nagao、T.Urushidani：“磷脂酰肌醇是参与胃质子泵送的 K 渗透性的重要决定因素”Am。