Development of a fluorescent monoclonal antibody-based assay to measure the allosteric effects of synthetic peptides on self-oligomerization of AGR2 protein
Identifikátory výsledku
Kód výsledku v 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>
Výsledek na webu
<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>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
Development of a fluorescent monoclonal antibody-based assay to measure the allosteric effects of synthetic peptides on self-oligomerization of AGR2 protein
Popis výsledku v původním jazyce
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
Název v anglickém jazyce
Development of a fluorescent monoclonal antibody-based assay to measure the allosteric effects of synthetic peptides on self-oligomerization of AGR2 protein
Popis výsledku anglicky
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
Klasifikace
Druh
J<sub>x</sub> - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)
CEP obor
EB - Genetika a molekulární biologie
OECD FORD obor
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Návaznosti výsledku
Projekt
Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.
Návaznosti
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Ostatní
Rok uplatnění
2013
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
Protein Science
ISSN
0961-8368
e-ISSN
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Svazek periodika
22
Číslo periodika v rámci svazku
9
Stát vydavatele periodika
US - Spojené státy americké
Počet stran výsledku
13
Strana od-do
1266-1278
Kód UT WoS článku
000323410100011
EID výsledku v databázi Scopus
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