Cell Biology

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Different Functionals Domain of SpastinM87V Affect Cellular Microtubule Cutting

Received: 14 January 2019    Accepted: 25 February 2019    Published: 20 March 2019
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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).

DOI 10.11648/j.cb.20190701.11
Published in Cell Biology (Volume 7, Issue 1, June 2019)
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), 2024. Published by Science Publishing Group

Keywords

SpastinM87V, Functional Domain, Protein Expression, Microtubule Cutting

References
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Author Information
  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

  • Department of Orthopedics, the First Affiliated Hospital of Jinan University, Guangzhou, China

Cite This Article
  • 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

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    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

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    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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  • @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://download.sciencepg.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}
    }
    

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  • 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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