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Related Experiment Video

Updated: Dec 7, 2025

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy
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EMPAS: Electron Microscopy Screening for Endogenous Protein Architectures.

Gijeong Kim1, Seongmin Jang1, Eunhye Lee1

  • 1Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea.

Molecules and Cells
|September 25, 2020
PubMed
Summary
This summary is machine-generated.

We developed a new top-down method, Electron Microscopy screening for endogenous Protein ArchitectureS (EMPAS), to study complex protein structures within cells. EMPAS revealed a novel spiral architecture and diverse molecular architectures across various organisms.

Keywords:
microscopyproteinscreeningstructure

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Area of Science:

  • Structural Biology
  • Cell Biology
  • Biochemistry

Background:

  • Proteins assemble into macromolecular complexes for biological functions.
  • Traditional structural biology uses bottom-up methods, which may not capture endogenous complexity.
  • Endogenous molecular architectures are diverse and complex, requiring new investigation approaches.

Purpose of the Study:

  • To introduce a novel top-down approach, Electron Microscopy screening for endogenous Protein ArchitectureS (EMPAS).
  • To investigate the diverse and complex landscape of endogenous macromolecular architectures in an unbiased manner.
  • To explore the utility of EMPAS across different biological systems.

Main Methods:

  • Development of Electron Microscopy screening for endogenous Protein ArchitectureS (EMPAS), a top-down screening method.
  • Application of EMPAS to discover and characterize endogenous protein architectures.
  • Screening of molecular architectures in human embryonic stem cells (hESCs), mouse brains, cyanobacteria, and plant leaves.

Main Results:

  • Discovery of a novel spiral architecture, identified as AdhE.
  • Revealed diverse repertoires of endogenous molecular architectures in various cell and tissue types.
  • Demonstrated the capability of EMPAS to unbiasedly investigate complex macromolecular structures.

Conclusions:

  • EMPAS provides a powerful top-down approach for unbiased investigation of endogenous macromolecular architectures.
  • The method successfully identified novel structures and revealed architectural diversity across species.
  • EMPAS has the potential to become a key tool for studying cellular structural biology.