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Biophysical Reviews

, Volume 11, Issue 2, pp 149–155 | Cite as

Advances in domain and subunit localization technology for electron microscopy

  • Zuben P. BrownEmail author
  • Junichi Takagi
Review
  • 113 Downloads

Abstract

The award of the 2017 Nobel Prize in chemistry, ‘for developing cryo-electron microscopy for the high-resolution structure determination of biomolecules in solution’, was recognition that this method, and electron microscopy more generally, represent powerful techniques in the scientific armamentarium for atomic level structural assessment. Technical advances in equipment, software, and sample preparation, have allowed for high-resolution structural determination of a range of complex biological machinery such that the position of individual atoms within these mega-structures can be determined. However, not all targets are amenable to attaining such high-resolution structures and some may only be resolved at so-called intermediate resolutions. In these cases, other tools are needed to correctly characterize the domain or subunit orientation and architecture. In this review, we will outline various methods that can provide additional information to help understand the macro-level organization of proteins/biomolecular complexes when high-resolution structural description is not available. In particular, we will discuss the recent development and use of a novel protein purification approach, known as the the PA tag/NZ-1 antibody system, which provides numberous beneficial properties, when used in electron microscopy experimentation.

Keywords

PA tag NZ-1 Domain mapping Electron microscopy 

Notes

Acknowledgements

Thank you to Joachim Frank for providing helpful comments on an early draft of this manuscript. This review was funded by National Institute of Health (NIH) R01 GM 29169 (to Joachim Frank) and Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Innovative Drug Discovery and Life Science Research (BINDS) funded by Japan Agency for Medical Research and Development (AMED) under Grant Number JP18am0101075 (to Junichi Takagi).

Compliance with ethical standards

Conflict of interest

Zuben P. Brown declares that he has no conflict of interest. Junichi Takagi 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 2019

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular BiophysicsColumbia UniversityNew YorkUSA
  2. 2.Laboratory of Protein Synthesis and Expression, Institute for Protein ResearchOsaka UniversityOsakaJapan

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