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
- Tanaka, Hidekazu; Takafuji, Kazuaki; Taguchi, Akihiko; Wiriyasermkul, Pattama; Ohgaki, Ryuichi; Nagamori, Shushi; Suh, Pann-Ghill; Kanai, Yoshikatsu
- Adhesion; Cadherin; Cytoskeleton; Hippocampus; Plasticity; Synapse
- Issue Date
- PERGAMON-ELSEVIER SCIENCE LTD
- NEUROCHEMISTRY INTERNATIONAL, v.61, no.2, pp.240 - 250
- 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 beta- and alpha-catenins, as well as F-actin-related membrane anchor proteins such as integrins bridged with alpha-actinin-4, and Na+/K+-ATPase bridged with spectrins. A close relative of beta-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 NPRAP/neurojungin/delta 2-catenin, but not p120ctn/delta 1-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 beta-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.
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