Synaptojanin forms two separate complexes in the nerve terminal - Interactions with endophilin and amphiphysin
Synaptojanin forms two separate complexes in the nerve terminal - Interactions with endophilin and amphiphysin
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DOI：
10.1074/jbc.272.43.27239
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发表时间：
1997-10-24
影响因子：
4.8
通讯作者：
McPherson, PS
中科院分区：
文献类型：
--
作者：
Micheva, KD;Kay, BK;McPherson, PS
内嗜肽是最近发现的含有src同源3结构域的蛋白，是synaptojanin的主要体外结合伙伴。为了进一步表征嗜内肽，我们产生了一种抗肽抗体。脑内嗜内蛋白丰富，免疫荧光分析显示突触末端有高浓度的这种蛋白，在那里它与突触蛋白共定位。体外结合实验表明，内啡肽通过其src同源3结构域与synaptojanin结合，免疫沉淀分析显示，内啡肽与synaptojanin在神经末梢稳定结合。两栖素I和两栖素II通过src同源结构域与动力蛋白和突触蛋白相互作用，其位点不同于亲内啡肽，免疫沉淀抗体揭示了第二种稳定复合物，包括动力蛋白和突触蛋白，但不包括亲内啡蛋白。这些数据表明突触联蛋白存在于神经末梢的两个独立复合物中，并支持亲内啡在突触联蛋白功能调节中的重要作用。
Endophilin is a recently discovered src homology 3 domain-containing protein that is a major in vitro binding partner for synaptojanin. To further characterize endophilin, we generated an antipeptide antibody. Endophilin is enriched in the brain, and immunofluorescence analysis reveals a high concentration of the protein in synaptic terminals, where it colocalizes with synaptojanin. In vitro binding assays demonstrate that endophilin binds through its src homology 3 domain to synaptojanin, and immunoprecipitation analysis with the antiendophilin antibody reveals that endophilin is stably associated with synaptojanin in the nerve terminal. Immunoprecipitation with an antibody against amphiphysin I and II, which interact through their src homology 3 domains with dynamin and synaptojanin at sites distinct from those for endophilin, reveals a second stable complex, which includes dynamin and synaptojanin but excludes endophilin. These data demonstrate that synaptojanin is present in two separate complexes in the nerve terminal and support an important role for endophilin in the regulation of synaptojanin function.