Improved cryo-EM reconstruction of sub-50 kDa complexes using 2D template matching.

Authors

Kexin Zhang, Timothy Grant, Nikolaus Grigorieff

Published in

eLife. Volume 15. Oct 26, 2026. Epub Oct 26, 2026.

Abstract

Visualizing the structures of small proteins and complexes has been a longstanding challenge in single-particle cryo-electron microscopy (cryo-EM). Some of these targets have been successfully resolved by binding to antibody fragments (Fabs) or fusing with external scaffolds to increase their size. Recent advances in conventional single-particle techniques have enabled the determination of an increasing number of structures smaller than 100 kDa, achieving resolutions relevant to drug research. Compared to X-ray crystallography, cryo-EM preserves the near-native states of biomolecules, can resolve structural heterogeneity, and has the potential to apply to a wide range of targets. In this work, we demonstrate that the alignment and reconstruction of small macromolecular complexes can be improved using high-resolution structures as priors, combined with 2D template matching. Using this method, we reconstructed a previously intractable ∼43 kDa protein kinase and improved the density of its ligand-binding site. Our theoretical analysis predicts that this method can further extend single-particle cryo-EM to important drug-binding complexes well below 50 kDa.

PMID:
42930231
Bibliographic data and abstract were imported from PubMed on 26 Oct 2026.

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