Microchip-Based Structure Determination of Disease-Relevant p53

• Published in: Analytical chemistry (Washington) Vol. 92; no. 23; pp. 15558 - 15564
• Main Authors: Solares, Maria J, Jonaid, G. M, Luqiu, William Y, Liang, Yanping, Evans, Madison C, Dearnaley, William J, Sheng, Zhi, Kelly, Deborah F
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01.12.2020
• Subjects: Cancer; Cell Line, Tumor; Chemistry; Disease; Electron microscopy; Human tissues; Humans; Microchip Analytical Procedures; Mutation; Nanotechnology; p53 Protein; Post-translation; Protein Multimerization; Protein Structure, Quaternary; Semiconductors; Tumor suppressor genes; Tumor Suppressor Protein p53 - chemistry; Tumor Suppressor Protein p53 - metabolism; Ubiquitination
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.0c03599

The tumor suppressor protein TP53 (p53) plays a multifaceted role in all cells of the human body. Mutations in the TP53 gene are often involved in cancer induction and disease progression. Despite its important role in health and development, structural information for p53 remains incomplete. Here, we present a microchip-based technology to facilitate structural studies of p53 assemblies derived from human cancer cells. These devices do not introduce foreign sequences to the p53 gene and maintain naturally occurring post-translational modifications. Using cryo-electron microscopy, structures for the p53 monomer (∼50 kDa) and tetramer (∼200 kDa) were resolved to ∼4.8 and ∼7 Å, respectively. These structures revealed new insights for flexible regions of p53 along with biologically relevant ubiquitination sites. Collectively, the convergence of nanotechnology tools and structural imaging builds a strong framework to understand the oncogenic impact of p53 in human tissues.