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Method Article
Deborah F. Kelly
Pennsylvania State University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-7341-7435
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Tumor suppressor protein TP53 (p53) plays a multi-faceted role in all cells of the
human body. Sadly, mutations in the TP53 gene are involved in nearly ~50% of tumors,
spurring erratic cell growth and disease progression. Until recently, structural information
for p53 remained incomplete and there are limited studies on native p53 produced in
human tumors. Here, we present a highly reproducible and effective protocol to extract,
enrich, and purify native p53 protein assemblies from cancer cells for downstream
structural studies. This method does not introduce purification tags into the p53 gene and
maintains naturally occurring modifications. In conjunction with cryo-Electron Microscopy
techniques, we determined new structures for p53 monomers (~50 kDa) and tetramers
(~200 kDa) at spatial resolutions of ~4.8 Å and ~7 Å, respectively.1 These models
revealed new insights for flexible regions of p53 along with biologically-relevant
ubiquitination sites. Combining biochemical and structural imaging protocols, we aim to
build a better understanding of native p53’s impact in cancer formation.
Keywords
p53, cancer, cryo-Electron Microscopy, microchips
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Associated Publications
Microchip-Based Structure Determination of Disease-Relevant p53
by Maria J. Solares, G. M. Jonaid, William Y. Luqiu, +5
Analytical Chemistry (30 October, 2020)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Deborah F. Kelly
Pennsylvania State University
Corresponding Author
ORCiD: https://orcid.org/0000-0002-7341-7435
Version History
CURRENT STATUS: Posted
Version 1
Posted 21 Jan, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Tumor suppressor protein TP53 (p53) plays a multi-faceted role in all cells of the
human body. Sadly, mutations in the TP53 gene are involved in nearly ~50% of tumors,
spurring erratic cell growth and disease progression. Until recently, structural information
for p53 remained incomplete and there are limited studies on native p53 produced in
human tumors. Here, we present a highly reproducible and effective protocol to extract,
enrich, and purify native p53 protein assemblies from cancer cells for downstream
structural studies. This method does not introduce purification tags into the p53 gene and
maintains naturally occurring modifications. In conjunction with cryo-Electron Microscopy
techniques, we determined new structures for p53 monomers (~50 kDa) and tetramers
(~200 kDa) at spatial resolutions of ~4.8 Å and ~7 Å, respectively.1 These models
revealed new insights for flexible regions of p53 along with biologically-relevant
ubiquitination sites. Combining biochemical and structural imaging protocols, we aim to
build a better understanding of native p53’s impact in cancer formation.
Figure 1
Figure 2
Loading...
Associated Publications
Microchip-Based Structure Determination of Disease-Relevant p53
by Maria J. Solares, G. M. Jonaid, William Y. Luqiu, +5
Analytical Chemistry (30 October, 2020)