The study for the function of synapsins in Synaptic vesicles and secretory granules

突触小泡和分泌颗粒中突触蛋白功能的研究

• 批准号: 13680837
• 负责人: HOSAKA Masahiro
• 金额: $ 2.3万
• 依托单位: Gunma University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 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/grant/KAKENHI-PROJECT-13680837/
• 关键词: Synapsin; Synaptic Vesicles; Chromogranin A; Secretogranin III; ATPase; Secretory Granules

A project of Synapsin in Synaptic vesiclesSynapsins constitute a family of synaptic vesicle proteins essential for regulating neurotransmitter release. Only two domains are conserved in all synapsins : a short N-terminal A domain with a single phosphorylation site for cAMP-dependent protein kinase (PKA) and CaM Kinase I, and a large central C domain that binds ATP and may be enzymatic. With this research grant, we tried to show that the C-domain of synapsins works as an ATPase and which unknown substrates co-laborate with synapsins. For these aims, we employed a yeast two-hybrid system with rat cDNA library. We are still analysing some candidate proteins.A project of Sorting Mechanisms to Secretory Granule in Neuroendocrine CellsChromogranin A (CgA) is transported restrictedly to regulated secretory granules in neuroendocrine cells. Because the binding-prone property of CgA with secretory proteins may be essential for its sorting to secretory granules, we screened its binding partner proteins using a yeast two-hybrid system. As results, we identified that SgIII binds to CgA and directs CgA to secretory granules by specific interactions in various endocrine cells.

突触小泡中突触蛋白的一个项目突触蛋白构成了调节神经递质释放所必需的突触小泡蛋白家族。所有突触蛋白中只有两个保守结构域：一个短的 N 端 A 结构域，具有 cAMP 依赖性蛋白激酶 (PKA) 和 CaM 激酶 I 的单个磷酸化位点，以及一个结合 ATP 并可能具有酶促性的大中央 C 结构域。通过这项研究资助，我们试图证明突触蛋白的 C 结构域作为 ATP 酶发挥作用，以及哪些未知底物与突触蛋白协同作用。为了实现这些目标，我们采用了带有大鼠 cDNA 文库的酵母双杂交系统。我们仍在分析一些候选蛋白。神经内分泌细胞分泌颗粒的分选机制项目嗜铬粒蛋白A（CgA）被限制性地转运到神经内分泌细胞中调节的分泌颗粒。由于 CgA 与分泌蛋白的易于结合的特性可能对其分选到分泌颗粒至关重要，因此我们使用酵母双杂交系统筛选了其结合伴侣蛋白。结果，我们发现 SgIII 与 CgA 结合，并通过各种内分泌细胞中的特异性相互作用将 CgA 引导至分泌颗粒。

项目成果

Hosaka M. Watanabe T, Sakai Y, Uchiyama Y, Takeuchi T.: "Identification of a chromogranin A domain that mediates binding to secretogranin III and targeting to secretory granules in pituitary cells and pancreatic β-cells"Mol Biol Cell.. 13. 3388-3399 (2002

Hosaka M. Watanabe T、Sakai Y、Uchiyama Y、Takeuchi T.：“介导与分泌粒蛋白 III 结合并靶向垂体细胞和胰腺 β 细胞中的分泌颗粒的嗜铬粒蛋白 A 结构域的鉴定”Mol Biol Cell.. 13。 3388-3399（2002年

Sakai Y, Hosaka M et al.: "Immunocytochemical localization of secretogranin III in the anterior lobe of male rat pituitary glands"J Histochem Cytochem. 51. 227-238 (2002)

Sakai Y、Hosaka M 等人：“雄性大鼠垂体前叶中促分泌素 III 的免疫细胞化学定位”J Histochem Cytochem。