Development of a fluorescent monoclonal antibody-based assay to measure the allosteric effects of synthetic peptides on self-oligomerization of AGR2 protein

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00209805%3A_____%2F13%3A%230000420" target="_blank" >RIV/00209805:_____/13:#0000420 - isvavai.cz</a>
Result on the web
<a href="http://onlinelibrary.wiley.com/doi/10.1002/pro.2299/abstract;jsessionid=F8B1804D27F005381295F370CA40BB77.f04t04" target="_blank" >http://onlinelibrary.wiley.com/doi/10.1002/pro.2299/abstract;jsessionid=F8B1804D27F005381295F370CA40BB77.f04t04</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1002/pro.2299" target="_blank" >10.1002/pro.2299</a>

Result language
angličtina
Original language name
Development of a fluorescent monoclonal antibody-based assay to measure the allosteric effects of synthetic peptides on self-oligomerization of AGR2 protein
Original language description
Many regulatory proteins are homo-oligomeric and designing assays that measure self-assembly will provide novel approaches to study protein allostery and screen for novel small molecule modulators of protein-interactions. We present an assay to begin todefine the biochemical determinants that regulate dimerization of the cancer-associated oncoprotein AGR2. A two site-sandwich microtiter assay (2S MTA) was designed using a DyLight800-labeled monoclonal antibody that binds to an epitope in AGR2 to screenfor synthetic self-peptides that might regulate dimer stability. Peptides derived from the intrinsically disordered N-terminal region of AGR2 increase in trans oligomer stability as defined using the 2S MTA assay. A DSS-crosslinking assay that traps theAGR2 dimer through K95-K95 adducts confirmed that ?45-AGR2 was a more stable dimer using denaturing gel electrophoresis. A titration of wt-AGR2, ?45-AGR2 (more stable dimer), and monomeric AGR2E60A revealed that ?45-AGR2 was more active
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EB - Genetics and molecular biology
OECD FORD branch
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Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year
2013
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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