Generation of stable microtubule superstructures by binding of peptide-fused tetrameric proteins to inside and outside

Microtubules play important roles in biological functions by forming superstructures, such as doublets and branched structures, in vivo. Despite the importance, it is challenging to construct these superstructures in vitro. Here, we designed a tetrameric fluorescent protein Azami-Green (AG) fused wi...

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Bibliographic Details
Published in:	Science advances Vol. 8; no. 36; p. eabq3817
Main Authors:	Inaba, Hiroshi, Sueki, Yurina, Ichikawa, Muneyoshi, Kabir, Arif Md. Rashedul, Iwasaki, Takashi, Shigematsu, Hideki, Kakugo, Akira, Sada, Kazuki, Tsukazaki, Tomoya, Matsuura, Kazunori
Format:	Journal Article
Language:	English
Published:	American Association for the Advancement of Science 09-09-2022
Subjects:	Biophysics Physical and Materials Sciences SciAdv r-articles Structural Biology
Online Access:	Get full text
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Summary:	Microtubules play important roles in biological functions by forming superstructures, such as doublets and branched structures, in vivo. Despite the importance, it is challenging to construct these superstructures in vitro. Here, we designed a tetrameric fluorescent protein Azami-Green (AG) fused with His-tag and Tau-derived peptide (TP), TP-AG, to generate the superstructures. Main binding sites of TP-AG can be controlled to the inside and outside of microtubules by changing the polymerization conditions. The binding of TP-AG to the inside promoted microtubule formation and generated rigid and stable microtubules. The binding of TP-AG to the outside induced various microtubule superstructures, including doublets, multiplets, branched structures, and extremely long microtubules by recruiting tubulins to microtubules. Motile microtubule aster structures were also constructed by TP-AG. The generation of various microtubule superstructures by a single type of exogenous protein is a new concept for understanding the functions of microtubules and constructing microtubule-based nanomaterials. Peptide-fused tetrameric proteins induce the generation of stable microtubule superstructures such as doublets and branches.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Present address: Japan Synchrotron Radiation Research Institute (JASRI), SPring-8, 1-1-1 Kouto, Sayo, Hyogo, 679-5148, Japan. These authors contributed equally to this work. Present address: State Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, Department of Biochemistry and Biophysics, School of Life Sciences, Fudan University, Shanghai 200438, China.
ISSN:	2375-2548 2375-2548
DOI:	10.1126/sciadv.abq3817