Počet záznamů: 1
Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation
- 1.
SYSNO ASEP 0534692 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Functional Assay to Correlate Protein Oligomerization States with Membrane Pore Formation Tvůrce(i) Šachl, Radek (UFCH-W) RID, ORCID
Čujová, Sabína (UFCH-W) ORCID
Singh, Vandana (UFCH-W)
Riegerová, Petra (UFCH-W) ORCID
Kapusta, Peter (UFCH-W) RID, ORCID
Müller, H.-M. (DE)
Steringer, J. P. (DE)
Hof, Martin (UFCH-W) RID, ORCID
Nickel, W. (DE)Zdroj.dok. Analytical Chemistry. - : American Chemical Society - ISSN 0003-2700
Roč. 92, č. 22 (2020), s. 14861-14866Poč.str. 6 s. Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova Peptides and proteins ; Oligomerization ; Fluorescence, ; Oligomers, Vědní obor RIV CF - Fyzikální chemie a teoretická chemie Obor OECD Physical chemistry CEP GC20-01401J GA ČR - Grantová agentura ČR GX19-26854X GA ČR - Grantová agentura ČR Způsob publikování Omezený přístup Institucionální podpora UFCH-W - RVO:61388955 UT WOS 000592852900002 EID SCOPUS 85096123765 DOI 10.1021/acs.analchem.0c03276 Anotace 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. Pracoviště Ústav fyzikální chemie J.Heyrovského Kontakt Michaela Knapová, michaela.knapova@jh-inst.cas.cz, Tel.: 266 053 196 Rok sběru 2021 Elektronická adresa http://hdl.handle.net/11104/0312867
Počet záznamů: 1