The research is aimed to explore the effect of different functional domains of SpastinM87V on microtubule cutting of Hela cells. First, the Spastin gene was extracted and its mutants were constructed and identified. Then, Spastin and its mutants were transfected into Hela cells and their cutting effects on microtubules were observed. The results showed that SpastinM87V and its truncated prokaryotic plasmids, including GST-SpastinN197, GST-Spastin△AAA, GST-Spastin△N1 and GST-Spastin△N2 were constructed, and the corresponding fusion proteins were expressed in vitro. In addition, SpastinM87V and its truncated eukaryotic plasmids including GFP-SpastinN197, GFP-Spastin△AAA, GFP-Spastin△N1 and GFP-Spastin△N2 were successfully constructed and introduced into Hela cells. At last, the microtubules of Hela cells could be cut into small fragments by SpastinM87V and Spastin△N1. Furthermore, the fluorescence intensity of the microtubules of Hela cells in the SpastinM87V and Spastin△N1 groups were significantly weaker than in the control group. In this way, Spastin is mainly involved in cell microtubule cutting, and the cutting function by SpastinM87V must contain complete microtubule binding- domain (MTBD) and cutting domain (AAA domain).
| Published in | Cell Biology (Volume 7, Issue 1) |
| DOI | 10.11648/j.cb.20190701.11 |
| Page(s) | 1-9 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2019. Published by Science Publishing Group |
SpastinM87V, Functional Domain, Protein Expression, Microtubule Cutting
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APA Style
Yang Yuhao, Zhang Guowei, Ji Zhisheng, Cai Zhenbin, Liu Qiuling, et al. (2019). Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting. Cell Biology, 7(1), 1-9. https://doi.org/10.11648/j.cb.20190701.11
ACS Style
Yang Yuhao; Zhang Guowei; Ji Zhisheng; Cai Zhenbin; Liu Qiuling, et al. Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting. Cell Biol. 2019, 7(1), 1-9. doi: 10.11648/j.cb.20190701.11
AMA Style
Yang Yuhao, Zhang Guowei, Ji Zhisheng, Cai Zhenbin, Liu Qiuling, et al. Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting. Cell Biol. 2019;7(1):1-9. doi: 10.11648/j.cb.20190701.11
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Download@article{10.11648/j.cb.20190701.11,
author = {Yang Yuhao and Zhang Guowei and Ji Zhisheng and Cai Zhenbin and Liu Qiuling and Li Shaojin and Lin Hongsheng},
title = {Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting},
journal = {Cell Biology},
volume = {7},
number = {1},
pages = {1-9},
doi = {10.11648/j.cb.20190701.11},
url = {https://doi.org/10.11648/j.cb.20190701.11},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.cb.20190701.11},
abstract = {The research is aimed to explore the effect of different functional domains of SpastinM87V on microtubule cutting of Hela cells. First, the Spastin gene was extracted and its mutants were constructed and identified. Then, Spastin and its mutants were transfected into Hela cells and their cutting effects on microtubules were observed. The results showed that SpastinM87V and its truncated prokaryotic plasmids, including GST-SpastinN197, GST-Spastin△AAA, GST-Spastin△N1 and GST-Spastin△N2 were constructed, and the corresponding fusion proteins were expressed in vitro. In addition, SpastinM87V and its truncated eukaryotic plasmids including GFP-SpastinN197, GFP-Spastin△AAA, GFP-Spastin△N1 and GFP-Spastin△N2 were successfully constructed and introduced into Hela cells. At last, the microtubules of Hela cells could be cut into small fragments by SpastinM87V and Spastin△N1. Furthermore, the fluorescence intensity of the microtubules of Hela cells in the SpastinM87V and Spastin△N1 groups were significantly weaker than in the control group. In this way, Spastin is mainly involved in cell microtubule cutting, and the cutting function by SpastinM87V must contain complete microtubule binding- domain (MTBD) and cutting domain (AAA domain).},
year = {2019}
}
TY - JOUR T1 - Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting AU - Yang Yuhao AU - Zhang Guowei AU - Ji Zhisheng AU - Cai Zhenbin AU - Liu Qiuling AU - Li Shaojin AU - Lin Hongsheng Y1 - 2019/03/20 PY - 2019 N1 - https://doi.org/10.11648/j.cb.20190701.11 DO - 10.11648/j.cb.20190701.11 T2 - Cell Biology JF - Cell Biology JO - Cell Biology SP - 1 EP - 9 PB - Science Publishing Group SN - 2330-0183 UR - https://doi.org/10.11648/j.cb.20190701.11 AB - The research is aimed to explore the effect of different functional domains of SpastinM87V on microtubule cutting of Hela cells. First, the Spastin gene was extracted and its mutants were constructed and identified. Then, Spastin and its mutants were transfected into Hela cells and their cutting effects on microtubules were observed. The results showed that SpastinM87V and its truncated prokaryotic plasmids, including GST-SpastinN197, GST-Spastin△AAA, GST-Spastin△N1 and GST-Spastin△N2 were constructed, and the corresponding fusion proteins were expressed in vitro. In addition, SpastinM87V and its truncated eukaryotic plasmids including GFP-SpastinN197, GFP-Spastin△AAA, GFP-Spastin△N1 and GFP-Spastin△N2 were successfully constructed and introduced into Hela cells. At last, the microtubules of Hela cells could be cut into small fragments by SpastinM87V and Spastin△N1. Furthermore, the fluorescence intensity of the microtubules of Hela cells in the SpastinM87V and Spastin△N1 groups were significantly weaker than in the control group. In this way, Spastin is mainly involved in cell microtubule cutting, and the cutting function by SpastinM87V must contain complete microtubule binding- domain (MTBD) and cutting domain (AAA domain). VL - 7 IS - 1 ER -
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