The fight against human disease requires a multidisciplinary scientific approach. Applying tools from seemingly unrelated areas, such as materials science and molecular biology, researchers can overcome long-standing challenges to improve knowledge of molecular pathologies. Here, custom-designed substrates composed of silicon nitride (SiN) are used to study the 3D attributes of tumor suppressor proteins that function in DNA repair events. New on-chip preparation strategies enable the isolation of native protein complexes from human cancer cells. Combined techniques of cryo-electron microscopy (EM) and molecular modeling reveal a new modified form of the p53 tumor suppressor present in aggressive glioblastoma multiforme cancer cells. Taken together, the findings provide a radical new design for cryo-EM substrates to evaluate the structures of disease-related macromolecules.

Supporting InformationThe Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.0c03599. Materials and methods and description for the supporting movies (PDF) EM structure of the p53 monomer with rotation (MOV) Cross sections through the p53 monomer structure (MOV) EM structure of the p53 tetramer with rotation (MOV) Cross sections through the p53 tetramer structure (MOV) Model for a putative p53 tetramer formation upon double-stranded DNA (MOV)

Interest in cryo-Electron Microscopy (EM) imaging has skyrocketed in recent years due to its pristine views of macromolecules and materials. As advances in instrumentation and computing algorithms spurred this progress,...