Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
The result's identifiers
Result code in 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>
Result on the web
<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>
Alternative languages
Result language
angličtina
Original language name
Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
Original language description
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.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10403 - Physical chemistry
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2020
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Analytical Chemistry
ISSN
0003-2700
e-ISSN
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Volume of the periodical
92
Issue of the periodical within the volume
22
Country of publishing house
US - UNITED STATES
Number of pages
6
Pages from-to
14861-14866
UT code for WoS article
000592852900002
EID of the result in the Scopus database
2-s2.0-85096123765