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首页> 外文期刊>Neurochemistry International: The International Journal for the Rapid Publication of Critical Reviews, Preliminary and Original Research Communications in Neurochemistry >Linkage of N-cadherin to multiple cytoskeletal elements revealed by a proteomic approach in hippocampal neurons
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Linkage of N-cadherin to multiple cytoskeletal elements revealed by a proteomic approach in hippocampal neurons

机译：蛋白质组学方法在海马神经元中揭示N-钙粘着蛋白与多种细胞骨架元素的联系

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The CNS synapse is an adhesive junction differentiated for chemical neurotransmission and is equipped with presynaptic vesicles and postsynaptic neurotransmitter receptors. Cell adhesion molecule cadherins not only maintain connections between pre- and postsynaptic membranes but also modulate the efficacy of synaptic transmission. Although the components of the cadherin-mediated adhesive apparatus have been studied extensively in various cell systems, the complete picture of these components, particularly at the synaptic junction, remains elusive. Here, we describe the proteomic assortment of the N-cadherin-mediated synaptic adhesion apparatus in cultured hippocampal neurons. N-cadherin immunoprecipitated from Triton X-100-solubilized neuronal extract contained equal amounts of β- and α-catenins, as well as F-actin-related membrane anchor proteins such as integrins bridged with α-actinin-4, and Na +/K +-ATPase bridged with spectrins. A close relative of β-catenin, plakoglobin, and its binding partner, desmoplakin, were also found, suggesting that a subset of the N-cadherin-mediated adhesive apparatus also anchors intermediate filaments. Moreover, dynein heavy chain and LEK1/CENPF/mitosin were found. This suggests that internalized pools of N-cadherin in trafficking vesicles are conveyed by dynein motors on microtubules. In addition, ARVCF and NPRAPeurojungin/ δ2-catenin, but not p120ctn/δ1-catenin or plakophilins-1, -2, -3, -4 (p0071), were found, suggesting other possible bridges to microtubules. Finally, synaptic stimulation by membrane depolarization resulted in an increased 93-kDa band, which corresponded to proteolytically truncated β-catenin. The integration of three different classes of cytoskeletal systems found in the synaptic N-cadherin complex may imply a dynamic switching of adhesive scaffolds in response to synaptic activity.
机译：中枢神经系统突触是为化学神经传递而分化的粘附连接，并配备有突触前囊泡和突触后神经递质受体。细胞粘附分子钙粘着蛋白不仅维持突触前膜和突触后膜之间的连接，而且调节突触传递的功效。尽管已经在各种细胞系统中广泛研究了钙粘蛋白介导的粘附装置的组件，但是这些组件的完整图片，尤其是在突触连接处，仍然难以捉摸。在这里，我们描述了在培养的海马神经元中N-钙黏着蛋白介导的突触粘附设备的蛋白质组学分类。从Triton X-100溶解的神经元提取物中免疫沉淀的N-钙粘着蛋白包含等量的β-和α-catenins，以及F-actin相关的膜锚蛋白，例如与α-actinin-4桥接的整合素和Na + / K + -ATPase与血影蛋白桥接。还发现了β-连环蛋白，珠蛋白和其结合伴侣desmoplakin的近亲，这表明N-钙粘蛋白介导的粘附装置的一部分也锚定了中间丝。此外，发现了动力蛋白重链和LEK1 / CENPF /丝裂霉素。这表明运输囊泡中的N-钙粘着蛋白的内在池是由微管上的动力蛋白驱动的。此外，还发现了ARVCF和NPRAP /神经氨酸/δ2-catenin，但未发现p120ctn /δ1-catenin或plakophilins-1，-2，-3，-4（p0071），这表明与微管的其他可能桥梁。最后，通过膜去极化的突触刺激导致增加的93 kDa条带，这对应于蛋白水解性截短的β-catenin。在突触N-钙黏着蛋白复合物中发现的三种不同类别的细胞骨架系统的整合可能暗示着粘性支架响应突触活性的动态转换。

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