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Biophysical Reviews

, Volume 10, Issue 2, pp 299–306 | Cite as

Multiple analyses of protein dynamics in solution

  • Tadayuki Ogawa
  • Nobutaka Hirokawa
Review

Abstract

The need for accurate description of protein behavior in solution has gained importance in various fields, including biophysics, biochemistry, structural biology, drug discovery, and antibody drugs. To achieve the desired accuracy, multiple precise analyses should be performed on the target molecule, compared, and effectively combined. This review focuses on the combination of multiple analyses in solution: size-exclusion chromatography (SEC), multi-angle light scattering (MALS), small-angle X-ray scattering (SAXS), analytical ultracentrifugation (AUC), and their complementary methods, such as atomic force microscopy (AFM) and mass spectrometry (MS). We also discuss the comparison between the determined molar mass value of not only the standard proteins, but of a target molecule tubulin and its depolymerizing protein, KIF2, as an example. The comparison of the estimated molar mass value from the different methods provides additional information about the target molecule, because the value reflects the dynamically changing states of the target molecule in solution. The combination and integration of multiple methods will permit a deeper understanding of protein dynamics in solution.

Keywords

Size-exclusion chromatography (SEC) Multi-angle light scattering (MALS) Small-angle X-ray scattering (SAXS) Analytical ultracentrifugation (AUC) Microtubule Kinesin 

Notes

Acknowledgements

We thank Dr. F. Arisaka for the valuable discussion about the AUC and MALS analyses. We thank Mr. E. Tsuruta, Mr. K. Kurono, Mr. K. Watanabe, and Mr. T. Tokai of Shoko Science for their help with the SEC-RI-MALS analysis. We thank Drs. S. Sakamoto, T. Nirasawa (Bruker Daltonics), and D. Higo (Thermo Fisher Scientific) for the valuable discussions about the MS analysis. We thank all members of N. Hirokawa’s laboratory for the valuable discussions and help. We thank Ms. H. Sato, Ms. H. Fukuda, Mr. T. Akamatsu, and Mr. N. Onouchi for the technical assistance. This work was supported by Grant-in-Aid for Specially Promoted Research (MEXT KAKENHI grant numbers 18002013 and 23000013) and for Scientific Research (S)(MEXT KAKENHI grant number 16H06372) by the Ministry of Education, Culture, Sports, Science and Technology of Japan to N.H. SAXS analyses were supported by the Platform for Drug Discovery, Informatics, and Structural Life Science (PDIS) from the Ministry of Education, Culture, Sports, Science and Technology of Japan to T.O. and N.H. (proposal number 2048). This work was partially supported by an unrestricted endowment from the Japan Electron Optics Laboratory (JEOL) and Carl Zeiss.

Author contributions

T.O. and N.H. conceived the project. T.O. performed the experiments. T.O. and N.H. discussed and wrote the paper.

Compliance with ethical standards

Conflict of interest

Tadayuki Ogawa declares that he has no conflict of interest. Nobutaka Hirokawa declares that he has no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  1. 1.Department of Cell Biology and Anatomy, Graduate School of MedicineUniversity of TokyoBunkyo-ku, TokyoJapan
  2. 2.Center of Excellence in Genome Medicine ResearchKing Abdulaziz UniversityJeddahSaudi Arabia

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