A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique

Mihoko Maruyama; Yuki Hayashi; Hiroshi Y. Yoshikawa; Shino Okada; Haruhiko Koizumi; Masaru Tachibana; Shigeru Sugiyama; Hiroaki Adachi; Hiroyoshi Matsumura; Tsuyoshi Inoue; Kazufumi Takano; Satoshi Murakami; Masashi Yoshimura; Yusuke Mori

doi:10.1016/j.jcrysgro.2015.11.008

A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique

Mihoko Maruyama, Yuki Hayashi, Hiroshi Y. Yoshikawa, Shino Okada, Haruhiko Koizumi, Masaru Tachibana, Shigeru Sugiyama, Hiroaki Adachi, Hiroyoshi Matsumura, Tsuyoshi Inoue, Kazufumi Takano, Satoshi Murakami, Masashi Yoshimura, Yusuke Mori

Materials Development Division

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

We developed a protein crystallization technique using a 0.0–2.0 w/v% agarose gel solution combined with a stirring technique for the purpose of controlling the crystal number in the gelled solutions. To confirm the stirring effect in the gelled solution, we investigated the nucleation probability and growth rate of the crystals produced using this method. The stirring operation by a rotary shaker affected the behavior of protein molecules in the gelled solution, and both a significant decrease in the nucleation rate and an enhancement of the crystal growth rate were achieved by the method. As a result, we concluded that the proposed technique, the stirring technique in a gel solution, was effective for generating protein crystals of sufficient and increased mechanical stability.

Original language	English
Pages (from-to)	172-178
Number of pages	7
Journal	Journal of Crystal Growth
Volume	452
DOIs	https://doi.org/10.1016/j.jcrysgro.2015.11.008
Publication status	Published - 2016 Oct 15

Keywords

A1. Fluid flows
A1. Nucleation
B1. Proteins
Crystallization technique
Nucleation control

ASJC Scopus subject areas

Condensed Matter Physics
Inorganic Chemistry
Materials Chemistry

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Maruyama, M., Hayashi, Y., Yoshikawa, H. Y., Okada, S., Koizumi, H., Tachibana, M., Sugiyama, S., Adachi, H., Matsumura, H., Inoue, T., Takano, K., Murakami, S., Yoshimura, M., & Mori, Y. (2016). A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique. Journal of Crystal Growth, 452, 172-178. https://doi.org/10.1016/j.jcrysgro.2015.11.008

A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique. / Maruyama, Mihoko; Hayashi, Yuki; Yoshikawa, Hiroshi Y.; Okada, Shino; Koizumi, Haruhiko; Tachibana, Masaru; Sugiyama, Shigeru; Adachi, Hiroaki; Matsumura, Hiroyoshi; Inoue, Tsuyoshi; Takano, Kazufumi; Murakami, Satoshi; Yoshimura, Masashi; Mori, Yusuke.

In: Journal of Crystal Growth, Vol. 452, 15.10.2016, p. 172-178.

Research output: Contribution to journal › Article › peer-review

Maruyama, M, Hayashi, Y, Yoshikawa, HY, Okada, S, Koizumi, H, Tachibana, M, Sugiyama, S, Adachi, H, Matsumura, H, Inoue, T, Takano, K, Murakami, S, Yoshimura, M & Mori, Y 2016, 'A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique', Journal of Crystal Growth, vol. 452, pp. 172-178. https://doi.org/10.1016/j.jcrysgro.2015.11.008

Maruyama, Mihoko ; Hayashi, Yuki ; Yoshikawa, Hiroshi Y. ; Okada, Shino ; Koizumi, Haruhiko ; Tachibana, Masaru ; Sugiyama, Shigeru ; Adachi, Hiroaki ; Matsumura, Hiroyoshi ; Inoue, Tsuyoshi ; Takano, Kazufumi ; Murakami, Satoshi ; Yoshimura, Masashi ; Mori, Yusuke. / A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique. In: Journal of Crystal Growth. 2016 ; Vol. 452. pp. 172-178.

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title = "A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique",

abstract = "We developed a protein crystallization technique using a 0.0–2.0 w/v% agarose gel solution combined with a stirring technique for the purpose of controlling the crystal number in the gelled solutions. To confirm the stirring effect in the gelled solution, we investigated the nucleation probability and growth rate of the crystals produced using this method. The stirring operation by a rotary shaker affected the behavior of protein molecules in the gelled solution, and both a significant decrease in the nucleation rate and an enhancement of the crystal growth rate were achieved by the method. As a result, we concluded that the proposed technique, the stirring technique in a gel solution, was effective for generating protein crystals of sufficient and increased mechanical stability.",

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author = "Mihoko Maruyama and Yuki Hayashi and Yoshikawa, {Hiroshi Y.} and Shino Okada and Haruhiko Koizumi and Masaru Tachibana and Shigeru Sugiyama and Hiroaki Adachi and Hiroyoshi Matsumura and Tsuyoshi Inoue and Kazufumi Takano and Satoshi Murakami and Masashi Yoshimura and Yusuke Mori",

note = "Funding Information: We acknowledge Mr. Ryota Mori for his experimental support. This work was supported in part by the Osaka University Program for the Support of Networking among Present and Future Researchers. We also acknowledge funding in the form of a JSAP KAKENHI Grant-in-Aid for Scientific Research B (Grant no. 23360011 ) to Y.M. Publisher Copyright: {\textcopyright} 2016 Elsevier B.V. Copyright: Copyright 2016 Elsevier B.V., All rights reserved.",

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doi = "10.1016/j.jcrysgro.2015.11.008",

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T1 - A crystallization technique for obtaining large protein crystals with increased mechanical stability using agarose gel combined with a stirring technique

AU - Maruyama, Mihoko

AU - Hayashi, Yuki

AU - Yoshikawa, Hiroshi Y.

AU - Okada, Shino

AU - Koizumi, Haruhiko

AU - Tachibana, Masaru

AU - Sugiyama, Shigeru

AU - Adachi, Hiroaki

AU - Matsumura, Hiroyoshi

AU - Inoue, Tsuyoshi

AU - Takano, Kazufumi

AU - Murakami, Satoshi

AU - Yoshimura, Masashi

AU - Mori, Yusuke

N1 - Funding Information: We acknowledge Mr. Ryota Mori for his experimental support. This work was supported in part by the Osaka University Program for the Support of Networking among Present and Future Researchers. We also acknowledge funding in the form of a JSAP KAKENHI Grant-in-Aid for Scientific Research B (Grant no. 23360011 ) to Y.M. Publisher Copyright: © 2016 Elsevier B.V. Copyright: Copyright 2016 Elsevier B.V., All rights reserved.

PY - 2016/10/15

Y1 - 2016/10/15

N2 - We developed a protein crystallization technique using a 0.0–2.0 w/v% agarose gel solution combined with a stirring technique for the purpose of controlling the crystal number in the gelled solutions. To confirm the stirring effect in the gelled solution, we investigated the nucleation probability and growth rate of the crystals produced using this method. The stirring operation by a rotary shaker affected the behavior of protein molecules in the gelled solution, and both a significant decrease in the nucleation rate and an enhancement of the crystal growth rate were achieved by the method. As a result, we concluded that the proposed technique, the stirring technique in a gel solution, was effective for generating protein crystals of sufficient and increased mechanical stability.

AB - We developed a protein crystallization technique using a 0.0–2.0 w/v% agarose gel solution combined with a stirring technique for the purpose of controlling the crystal number in the gelled solutions. To confirm the stirring effect in the gelled solution, we investigated the nucleation probability and growth rate of the crystals produced using this method. The stirring operation by a rotary shaker affected the behavior of protein molecules in the gelled solution, and both a significant decrease in the nucleation rate and an enhancement of the crystal growth rate were achieved by the method. As a result, we concluded that the proposed technique, the stirring technique in a gel solution, was effective for generating protein crystals of sufficient and increased mechanical stability.

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KW - A1. Nucleation

KW - B1. Proteins

KW - Crystallization technique

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