Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
Identifikátory výsledku
Kód výsledku v IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388955%3A_____%2F20%3A00534692" target="_blank" >RIV/61388955:_____/20:00534692 - isvavai.cz</a>
Výsledek na webu
<a href="http://hdl.handle.net/11104/0312867" target="_blank" >http://hdl.handle.net/11104/0312867</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/acs.analchem.0c03276" target="_blank" >10.1021/acs.analchem.0c03276</a>
Alternativní jazyky
Jazyk výsledku
angličtina
Název v původním jazyce
Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
Popis výsledku v původním jazyce
In-membrane oligomerization is decisive for the function (or dysfunction) of many proteins. Techniques were developed to characterize membrane-inserted oligomers and the hereby obtained oligomerization states were intuitively related to the function of these proteins. However, in many cases, it is unclear whether the obtained oligomerization states are functionally relevant or are merely the consequence of nonspecific aggregation. Using fibroblast growth factor 2 (FGF2) as a model system, we addressed this methodological challenge. FGF2 oligomerizes in a PI(4,5)P2-dependent manner at the inner plasma membrane leaflet. This process results in membrane insertion and the formation of a lipidic membrane pore, the key intermediate in unconventional secretion of FGF2. To tackle the problem of discriminating functional oligomers from irrelevant aggregates, we present a statistical single molecule and single vesicle assay determining the brightness of individually diffusing in-membrane oligomers and correlating their oligomerization state with membrane pore formation. Importantly, time-dependent membrane pore formation was analyzed with an ensemble of single vesicles providing detailed statistics. Our findings demonstrate that quantifying oligomeric states alone does not allow for a deep understanding of the structure–function relationship of membrane-inserted oligomers.
Název v anglickém jazyce
Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
Popis výsledku anglicky
In-membrane oligomerization is decisive for the function (or dysfunction) of many proteins. Techniques were developed to characterize membrane-inserted oligomers and the hereby obtained oligomerization states were intuitively related to the function of these proteins. However, in many cases, it is unclear whether the obtained oligomerization states are functionally relevant or are merely the consequence of nonspecific aggregation. Using fibroblast growth factor 2 (FGF2) as a model system, we addressed this methodological challenge. FGF2 oligomerizes in a PI(4,5)P2-dependent manner at the inner plasma membrane leaflet. This process results in membrane insertion and the formation of a lipidic membrane pore, the key intermediate in unconventional secretion of FGF2. To tackle the problem of discriminating functional oligomers from irrelevant aggregates, we present a statistical single molecule and single vesicle assay determining the brightness of individually diffusing in-membrane oligomers and correlating their oligomerization state with membrane pore formation. Importantly, time-dependent membrane pore formation was analyzed with an ensemble of single vesicles providing detailed statistics. Our findings demonstrate that quantifying oligomeric states alone does not allow for a deep understanding of the structure–function relationship of membrane-inserted oligomers.
Klasifikace
Druh
J<sub>imp</sub> - Článek v periodiku v databázi Web of Science
CEP obor
—
OECD FORD obor
10403 - Physical chemistry
Návaznosti výsledku
Projekt
Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.
Návaznosti
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Ostatní
Rok uplatnění
2020
Kód důvěrnosti údajů
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Údaje specifické pro druh výsledku
Název periodika
Analytical Chemistry
ISSN
0003-2700
e-ISSN
—
Svazek periodika
92
Číslo periodika v rámci svazku
22
Stát vydavatele periodika
US - Spojené státy americké
Počet stran výsledku
6
Strana od-do
14861-14866
Kód UT WoS článku
000592852900002
EID výsledku v databázi Scopus
2-s2.0-85096123765