Crystallization and preliminary X-ray crystallographic study of a 3.8-MDa respiratory supermolecule hemocyanin

Asuka Matsuno, Zuoqi Gai, Miyuki Tanaka, Koji Kato, Sanae Kato, Tsuyoshi Katoh, Takeshi Shimizu, Takeya Yoshioka, Hideki Kishimura, Yoshikazu Tanaka, Min Yao

研究成果: Article

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Many molluscs transport oxygen using a very large cylindrical multimeric copper-containing protein named hemocyanin. The molluscan hemocyanin forms a decamer (cephalopods) or multidecamer (gastropods) of approximately 330-450. kDa subunits, resulting in a molecular mass >3.3. MDa. Therefore, molluscan hemocyanin is one of the largest proteins. The reason why these organisms use such a large supermolecule for oxygen transport remains unclear. Atomic-resolution X-ray crystallographic analysis is necessary to unveil the detailed molecular structure of this mysterious large molecule. However, its propensity to dissociate in solution has hampered the crystallization of its intact form. In the present study, we successfully obtained the first crystals of an intact decameric molluscan hemocyanin. The diffraction dataset at 3.0-Å resolution was collected by merging the datasets of two isomorphic crystals. Electron microscopy analysis of the dissolved crystals revealed cylindrical particles. Furthermore, self-rotation function analysis clearly showed the presence of a fivefold symmetry with several twofold symmetries perpendicular to the fivefold axis. The absorption spectrum of the crystals showed an absorption peak around 345. nm. These results indicated that the crystals contain intact hemocyanin decamers in the oxygen-bound form.

元の言語English
ページ(範囲)379-382
ページ数4
ジャーナルJournal of Structural Biology
190
発行部数3
DOI
出版物ステータスPublished - 2015 6 1
外部発表Yes

ASJC Scopus subject areas

  • Structural Biology

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    Matsuno, A., Gai, Z., Tanaka, M., Kato, K., Kato, S., Katoh, T., Shimizu, T., Yoshioka, T., Kishimura, H., Tanaka, Y., & Yao, M. (2015). Crystallization and preliminary X-ray crystallographic study of a 3.8-MDa respiratory supermolecule hemocyanin. Journal of Structural Biology, 190(3), 379-382. https://doi.org/10.1016/j.jsb.2015.04.015