Electron cryomicroscopy as a powerful tool in biomedical research

Abstract

A human cell is a precisely regulated system that relies on the complex interaction of molecules. Structural insights into the cellular machinery at the atomic level allow us to understand the underlying regulatory mechanism and provide us with a roadmap for the development of novel drugs to fight diseases. Facilitated by recent technological breakthroughs, the Nobel prize-winning technique electron cryomicroscopy (cryo-EM) has become a versatile and extremely powerful tool to solve routinely near-atomic resolution three-dimensional protein structures. Consequently, it has become the focus of attention for structure-based drug design. In this review, we describe the basics of cryo-EM and highlight its growing role in biomedical research. Furthermore, we discuss latest developments as well as future perspectives.

Structure and Function of the γ-Secretase Complex

“…Recent advances in cryo-EM-particularly the development of direct electron detection in combination with computational single-particle analysis, image sorting, and threedimensional (3D) reconstruction-have revolutionized structural biology, providing atomicresolution structures of previously inaccessible protein complexes. 117 The size, complexity, hydrophobicity, and heterogeneous glycosylation of γ-secretase make it highly challenging to crystallize for X-ray diffraction. Other I-CLiP family members have proven to be amenable to crystallography, [118][119][120][121] but the rhomboid serine proteases, the S2P metalloproteases, and the presenilin homologue aspartyl proteases are much smaller and single polypeptides, with chemically and thermally stable orthologs found in the microbial world.…”

A comparative study of single-particle cryo-EM with liquid-nitrogen and liquid-helium cooling

“…Electron microscopy (EM) has become a key technique for determining the structures of biological macromolecules at high resolution (Vinothkumar & Henderson, 2016;Quentin & Raunser, 2018;Cheng, 2015;Kü hlbrandt, 2014a;Bai et al, 2015;Scapin et al, 2018), an essential step in understanding biological processes at the molecular level. The most fundamental and ultimately insurmountable limitation of the method is radiation damage to the sample.…”

Challenges and limitations in the studies of glycoproteins: A computational chemist's perspective

“…Due to numerous experimental challenges, molecular dynamics Electron microscopy (cryo-EM) has become an extremely powerful and versatile tool for the studies of glycoprotein structures at near-atomic resolution. 92 However, its potential has been limited by the glycoprotein structure minimum size it can be used to study and by image resolution. Technological advances, such as direct electron detector development and development of computational image analysis techniques currently are improving cryo-EM in the highresolution studies of large glycoprotein assemblies.…”

“…Electron microscopy (cryo‐EM) has become an extremely powerful and versatile tool for the studies of glycoprotein structures at near‐atomic resolution 92 . However, its potential has been limited by the glycoprotein structure minimum size it can be used to study and by image resolution.…”

“…The structures of the HCoV‐229E spike in the prefusion state measured by cryo‐EM. Reproduced with permission Reference 92. Copyright, 2021, Nat Commun…”