Technical approaches of 3D reconstruction from protein complex using the mixture of differently stained images: providing suggestive evidence for improving its resolution.

ABSTRACT: Negative staining electron microscopy remains a valuable tool for structural biology, particularly for initial characterization of large protein complexes. However, the limitations of single staining methods often result in incomplete structural information. Here, we present a novel multi-stain approach for negative staining electron microscopy, applied to the structural analysis of the pyruvate dehydrogenase E2 (PDH E2) complex. By integrating data from three distinct staining agents (uranyl acetate, ammonium phosphotungstate, and ammonium molybdate) we demonstrate significant improvements in structural resolution and detail. Our method improved the resolution from a range of approximately 27-31 Å (observed with individual stains) to about 21.7 Å in the combined dataset. This enhancement

SUBMITTER: Park YH 

PROVIDER: S-EPMC12119436 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature