Botulinum protease-cleaved SNARE fragments induce cytotoxicity in neuroblastoma cells.
Botulinum protease-cleaved SNARE fragments induce cytotoxicity in neuroblastoma cells.
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DOI：
10.1111/jnc.12645
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发表时间：
2014-06
影响因子：
4.7
通讯作者：
Davletov B
中科院分区：
文献类型：
--
作者：
Arsenault J;Cuijpers SA;Ferrari E;Niranjan D;Rust A;Leese C;O'Brien JA;Binz T;Davletov B
可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体（SNARE）对于胞吐、运输和神经突生长至关重要，其中囊泡SNARE针对其伴侣靶SNARE：25 kDa的突触体相关蛋白和突触融合蛋白。SNARE蛋白通常是膜结合的，但可以被肉毒杆菌神经毒素切割和释放。我们发现，肉毒杆菌蛋白酶C型和D型可以很容易地转导到内分泌细胞使用DNA转染试剂。在将C和D蛋白酶给予正常难治性Neuro 2A神经母细胞瘤细胞后，SNARE蛋白在数小时内被高效切割。值得注意的是，肉毒杆菌蛋白酶暴露导致细胞毒性证明了分光光度测定和碘化丙啶渗透到细胞核。直接将SNARE片段递送到神经母细胞瘤细胞中降低了与肉毒杆菌蛋白酶的应用类似的活力。我们观察到肉毒杆菌蛋白酶的协同细胞毒性作用，这可以通过可溶性SNARE片段的释放和相互作用来解释。我们首次表明，先前观察到的肉毒杆菌神经毒素/C在神经元中的细胞毒性可以在神经内分泌来源的细胞中实现，这对肉毒杆菌制剂的医学用途具有影响。
Soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs) are crucial for exocytosis, trafficking, and neurite outgrowth, where vesicular SNAREs are directed toward their partner target SNAREs: synaptosomal-associated protein of 25 kDa and syntaxin. SNARE proteins are normally membrane bound, but can be cleaved and released by botulinum neurotoxins. We found that botulinum proteases types C and D can easily be transduced into endocrine cells using DNA-transfection reagents. Following administration of the C and D proteases into normally refractory Neuro2A neuroblastoma cells, the SNARE proteins were cleaved with high efficiency within hours. Remarkably, botulinum protease exposures led to cytotoxicity evidenced by spectrophotometric assays and propidium iodide penetration into the nuclei. Direct delivery of SNARE fragments into the neuroblastoma cells reduced viability similar to botulinum proteases' application. We observed synergistic cytotoxic effects of the botulinum proteases, which may be explained by the release and interaction of soluble SNARE fragments. We show for the first time that previously observed cytotoxicity of botulinum neurotoxins/C in neurons could be achieved in cells of neuroendocrine origin with implications for medical uses of botulinum preparations.
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影响因子：
56.9
作者：
Dong, M;Yeh, F;Chapman, ER
通讯作者：
Chapman, ER
DOI：
10.1083/jcb.200406126
发表时间：
2005-02-14
期刊：
The Journal of cell biology
影响因子：
--
作者：
Berliocchi L;Fava E;Leist M;Horvat V;Dinsdale D;Read D;Nicotera P
通讯作者：
Nicotera P
影响因子：
9.9
作者：
Erbguth, FJ;Naumann, M
通讯作者：
Naumann, M
DOI：
10.1073/pnas.0903111106
发表时间：
2009-06-09
影响因子：
11.1
作者：
Chen, Sheng;Barbieri, Joseph T.
通讯作者：
Barbieri, Joseph T.
DOI：
10.1126/science.1224492
发表时间：
2012-09-14
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Gao Y;Zorman S;Gundersen G;Xi Z;Ma L;Sirinakis G;Rothman JE;Zhang Y
通讯作者：
Zhang Y