The Advantage of Small-angle Neutron Scattering Revealed through Analyzing the Overall Structure of a Fully Assembled Complex in Circadian Clock

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  • Yunoki Yasuhiro
    Institute for Integrated Radiation and Nuclear Science, Kyoto University
  • Matsumoto Atsushi
    National Institutes for Quantum Science and Technology
  • Morishima Ken
    Institute for Integrated Radiation and Nuclear Science, Kyoto University
  • Martel Anne
    Institut Laue-Langevin
  • Porcar Lionel
    Institut Laue-Langevin
  • Sato Nobuhiro
    Institute for Integrated Radiation and Nuclear Science, Kyoto University
  • Yogo Rina
    Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences Institute for Molecular Science (IMS), National Institutes of Natural Sciences Graduate School of Pharmaceutical Sciences, Nagoya City University
  • Tominaga Taiki
    Neutron Science and Technology Center, Comprehensive Research Organization for Science and Society
  • Yagi-Utsumi Maho
    Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences Institute for Molecular Science (IMS), National Institutes of Natural Sciences Graduate School of Pharmaceutical Sciences, Nagoya City University
  • Inoue Rintaro
    Institute for Integrated Radiation and Nuclear Science, Kyoto University
  • Kono Hidetoshi
    National Institutes for Quantum Science and Technology
  • Yagi Hirokazu
    Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences Graduate School of Pharmaceutical Sciences, Nagoya City University
  • Kato Koichi
    Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences Institute for Molecular Science (IMS), National Institutes of Natural Sciences Graduate School of Pharmaceutical Sciences, Nagoya City University
  • Sugiyama Masaaki
    Institute for Integrated Radiation and Nuclear Science, Kyoto University

Bibliographic Information

Other Title
  • 時計タンパク質複合体の構造解析を通して明らかとなった中性子小角散乱法の強み
  • トケイ タンパクシツ フクゴウタイ ノ コウゾウ カイセキ オ トオシテ アキラカ ト ナッタ チュウセイシ ショウカク サンランホウ ノ ツヨミ

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Abstract

<p>Cyanobacterial clock proteins; KaiA, KaiB and KaiC are periodically assembled into the large complex (ABC complex) in a 24-hour period in vitro. A cryo-EM study reported the structure of ABC complex. However, the locations of N-terminal domains of KaiA (AN domains) have not been resolved due to their high flexibility. For a better understanding of the assembly mechanisms, we investigated the overall structural model by integrating structural modelling based on the cryo-EM structure and small-angle X-ray/neutron scattering (SAXS/SANS) measurements. From the profiles of size exclusion chromatography (SEC)-SAXS measurements, the structural candidates of ABC complex were classified into three types (Type 1 ~ Type 3) from the viewpoint of the coordinate structure of AN domains. In order to refine the classified structures by the spatial arrangement of AN domains, we applied the SEC-inverse contrast matching (iCM)-SANS measurement to selectively observe KaiA in ABC complex which was prepared with hydrogenated KaiA along with 75%-deuterated KaiB and KaiC. The scattering from the latter components diminished and only the former component was visible in 100% D2O. By comparing the calculated SEC-iCM-SANS profiles for classified models and the experimental data, Type I were clearly the best structural model. In this report, we show the advantage of SEC-iCM-SANS measurement to resolve a large complex harboring dynamically fluctuating domains.</p>

Journal

  • hamon

    hamon 32 (4), 158-164, 2022-11-10

    The Japanese Society for Neutron Science

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