A Protein Interaction Node at the Neurotransmitter Release Site: Domains of Aczonin/Piccolo, Bassoon, CAST, and Rim Converge on the N-Terminal Domain of Munc13-1
A Protein Interaction Node at the Neurotransmitter Release Site: Domains of Aczonin/Piccolo, Bassoon, CAST, and Rim Converge on the N-Terminal Domain of Munc13-1
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DOI：
10.1523/jneurosci.1255-09.2009
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发表时间：
2009-10
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通讯作者：
Xiaolu Wang;Bin Hu;Agata Zięba;Nicole G. Neumann;M. Kasper-Sonnenberg;Annegret Honsbein;G. Hultqvist;Tim Conze;W. Witt;C. Limbach;M. Geitmann;H. Danielson;Richard Kolarow;G. Niemann;V. Lessmann;M. Kilimann
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作者：
Xiaolu Wang;Bin Hu;Agata Zięba;Nicole G. Neumann;M. Kasper-Sonnenberg;Annegret Honsbein;G. Hultqvist;Tim Conze;W. Witt;C. Limbach;M. Geitmann;H. Danielson;Richard Kolarow;G. Niemann;V. Lessmann;M. Kilimann
多结构域支架蛋白在突触发生和突触活动过程中组织神经递质囊泡动力学的分子机制。我们发现五个活性区蛋白的结构域集中在Munc13-1的n端区域的一个相互作用节点上，包括Rim1的锌指结构域、Bassoon的c端区域、CAST1/ELKS2的一个片段以及Aczonin/Piccolo或Bassoon的第三个卷曲结构域（CC3）。这种多结构域复合物可能构成了活性区蛋白质机制物理和功能整合的中心。azonin和巴松管之间的另一个连接是由azonin的第二个卷曲结构域介导的。重组Aczonin-CC3作为绿色荧光蛋白融合蛋白在培养的神经元中表达，以突触为靶点，抑制囊泡更新，提示其参与突触组装和活性。我们的研究结果表明，Aczonin、Bassoon、CAST1、Munc13和Rim是紧密且多重相互关联的，这表明Aczonin- cc3可以积极参与神经递质囊泡动力学，他们强调了Munc13-1的n端区域作为蛋白质相互作用的枢纽，通过增加三个新的结合伙伴来控制突触囊泡启动的机制潜力。
Multidomain scaffolding proteins organize the molecular machinery of neurotransmitter vesicle dynamics during synaptogenesis and synaptic activity. We find that domains of five active zone proteins converge on an interaction node that centers on the N-terminal region of Munc13-1 and includes the zinc-finger domain of Rim1, the C-terminal region of Bassoon, a segment of CAST1/ELKS2, and the third coiled-coil domain (CC3) of either Aczonin/Piccolo or Bassoon. This multidomain complex may constitute a center for the physical and functional integration of the protein machinery at the active zone. An additional connection between Aczonin and Bassoon is mediated by the second coiled-coil domain of Aczonin. Recombinant Aczonin-CC3, expressed in cultured neurons as a green fluorescent protein fusion protein, is targeted to synapses and suppresses vesicle turnover, suggesting involvements in synaptic assembly as well as activity. Our findings show that Aczonin, Bassoon, CAST1, Munc13, and Rim are closely and multiply interconnected, they indicate that Aczonin-CC3 can actively participate in neurotransmitter vesicle dynamics, and they highlight the N-terminal region of Munc13-1 as a hub of protein interactions by adding three new binding partners to its mechanistic potential in the control of synaptic vesicle priming.