Pregled bibliografske jedinice broj: 882266
Quantitative analysis of endocytic recycling of membrane proteins by monoclonal antibody-based recycling assays
Quantitative analysis of endocytic recycling of membrane proteins by monoclonal antibody-based recycling assays // Journal of cellular physiology, 232 (2017), 3; 463-476 doi:10.1002/jcp.25503 (međunarodna recenzija, članak, znanstveni)
CROSBI ID: 882266 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Quantitative analysis of endocytic recycling of membrane proteins by monoclonal antibody-based recycling assays
Autori
Blagojević Zagorac, Gordana ; Mahmutefendić, Hana ; Maćešić, Senka ; Karleuša, Ljerka ; Lučin, Pero
Izvornik
Journal of cellular physiology (0021-9541) 232
(2017), 3;
463-476
Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni
Ključne riječi
Endosomal recycling ; Recycling protocols ; Monoclonal antibodies ; MHC Class I recycling ; Transferrin receptor recycling
Sažetak
In this report, we present an analysis of several recycling protocols based on labeling of membrane proteins with specific monoclonal antibodies (mAbs). We analyzed recycling of membrane proteins that are internalized by clathrin-dependent endocytosis, represented by the transferrin receptor, and by clathrin-independent endocytosis, represented by the Major Histocompatibility Class I molecules. Cell surface membrane proteins were labeled with mAbs and recycling of mAb:protein complexes was determined by several approaches. Our study demonstrates that direct and indirect detection of recycled mAb:protein complexes at the cell surface underestimate the recycling pool, especially for clathrin-dependent membrane proteins that are rapidly reinternalized after recycling. Recycling protocols based on the capture of recycled mAb:protein complexes require the use of the Alexa Fluor 488 conjugated secondary antibodies or FITC-conjugated secondary antibodies in combination with inhibitors of endosomal acidification and degradation. Finally, protocols based on the capture of recycled proteins that are labeled with Alexa Fluor 488 conjugated primary antibodies and quenching of fluorescence by the anti-Alexa Fluor 488 displayed the same quantitative assessment of recycling as the antibody-capture protocols.
Izvorni jezik
Engleski
Znanstvena područja
Temeljne medicinske znanosti
POVEZANOST RADA
Projekti:
HRZZ-IP-2014-09-9564 - Karakterizacija preuređivanja endosomalnih membrana tijekom rane faze infekcije mišjim citomegalovirusom koje vodi k formiranju kompleksa za stvaranje viriona (AsCo) (Lučin, Pero) ( CroRIS)
Ustanove:
Medicinski fakultet, Rijeka
Profili:
Gordana Blagojević Zagorac
(autor)
Ljerka Karleuša
(autor)
Hana Mahmutefendić Lučin
(autor)
Pero Lučin
(autor)
Senka Maćešić
(autor)
Citiraj ovu publikaciju:
Časopis indeksira:
- Current Contents Connect (CCC)
- Web of Science Core Collection (WoSCC)
- Science Citation Index Expanded (SCI-EXP)
- SCI-EXP, SSCI i/ili A&HCI
- Scopus
- MEDLINE