Cryo‐EM‐On‐a‐Chip: Custom‐Designed Substrates for the 3D Analysis of Macromolecules

• Published in: Small (Weinheim an der Bergstrasse, Germany) Vol. 15; no. 21; pp. e1900918 - n/a
• Main Authors: Alden, Nick A., Varano, A. Cameron, Dearnaley, William J., Solares, Maria J., Luqiu, William Y., Liang, Yanping, Sheng, Zhi, McDonald, Sarah M., Damiano, John, McConnell, Jennifer, Dukes, Madeline J., Kelly, Deborah F.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.05.2019
• Subjects: breast cancer; Cancer; cryo‐electron microscopy; glioblastoma multiforme; Macromolecules; Materials science; Molecular biology; Nanotechnology; p53; Proteins; Silicon nitride; Silicon substrates; Tumors; glioblastoma multiforme; breast cancer; silicon nitride; cryo-electron microscopy; p53
• ISSN: 1613-6810; 1613-6829
• DOI: 10.1002/smll.201900918

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. Interest in the cryo‐electron microscopy (EM) field has recently skyrocketed, presenting new opportunities to study molecular systems. Here, new cryo‐EM techniques are developed through the use of custom‐designed substrates (Cryo‐Chips). These substrates enable structural studies of native tumor suppressors that are mutated in human cancer. Cryo‐Chip may be broadly used to study entities for both materials and life sciences applications.