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Pregled bibliografske jedinice broj: 603794

Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells

von Einem, Bjorn; Weber, Petra; Wagner, Michael; Malnar, Martina; Košiček, Marko; Hećimović, Silva; von Arnim, Christine; Schneckenburger, Herbert
Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells // International journal of molecular sciences, 13 (2012), 12; 15801-15812 doi:10.3390/ijms131215801 (međunarodna recenzija, članak, znanstveni)

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Naslov
Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells

Autori
Von Einem, Bjorn ; Weber, Petra ; Wagner, Michael ; Malnar, Martina ; Košiček, Marko ; Hećimović, Silva ; von Arnim, Christine ; Schneckenburger, Herbert

Izvornik
International journal of molecular sciences (1422-0067) 13 (2012), 12; 15801-15812

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
FRET ; cholesterol ; APP ; BACE1 ; NPC ; TIRFM ; neurodegeneration ; Laurdan ; generalized polarization

Sažetak
Förster resonance energy transfer (FRET) based techniques have recently been applied to study the interactions between β-site APP-cleaving enzyme-GFP (BACE1-GFP) and amyloid precursor protein-mRFP (APP-mRFP) in U373 glioblastoma cells. In this context the role of APP-BACE1 proximity in Alzheimer’s disease (AD) pathogenesis has been discussed. FRET was found to depend on intracellular cholesterol levels and associated alterations in membrane stiffness. Here, NPC1 null cells (CHO-NPC1-/-), exhibiting increased cholesterol levels and disturbed cholesterol transport similar to that observed in Niemann-Pick type C disease (NPC), were used to analyze the influence of altered cholesterol levels on APP-BACE1 proximity. Fluorescence lifetime measurements of whole CHO-wild type (WT) and CHO-NPC1-/- cells (EPI-illumination microscopy) as well as their plasma membranes (total internal reflection fluorescence microscopy, TIRFM) were performed. Additionally, generalized polarization (GP) measurements of CHO-WT and CHO-NPC1-/- cells incubated with the fluorescence marker laurdan were performed to determine membrane stiffness of plasma- and intracellular membranes. CHO-NPC1-/- cells showed higher membrane stiffness at intracellular- but not plasma membranes, equivalent to cholesterol accumulation in late endosomes/lysosomes. Along with higher membrane stiffness, the FRET efficiency between BACE1-GFP and APP-mRFP was reduced at intracellular membranes, but not within the plasma membrane of CHO-NPC1-/- . Our data show that FRET combined with TIRF is a powerful technique to determine protein proximity and membrane fluidity in cellular models of neurodegenerative diseases.

Izvorni jezik
Engleski

Znanstvena područja
Biologija



POVEZANOST RADA

Projekti:
098-0982522-2525 - Mehanizam djelovanja kolesterola u nastanku Alzheimerove bolesti (Katušić Hećimović, Silva, MZOS ) ( CroRIS)

Ustanove:
Institut "Ruđer Bošković", Zagreb

Profili:

Avatar Url Martina Malnar (autor)

Avatar Url Marko Košiček (autor)

Avatar Url Silva Katušić Hećimović (autor)

Poveznice na cjeloviti tekst rada:

doi doi.org fulir.irb.hr

Citiraj ovu publikaciju:

von Einem, Bjorn; Weber, Petra; Wagner, Michael; Malnar, Martina; Košiček, Marko; Hećimović, Silva; von Arnim, Christine; Schneckenburger, Herbert
Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells // International journal of molecular sciences, 13 (2012), 12; 15801-15812 doi:10.3390/ijms131215801 (međunarodna recenzija, članak, znanstveni)
Von Einem, B., Weber, P., Wagner, M., Malnar, M., Košiček, M., Hećimović, S., von Arnim, C. & Schneckenburger, H. (2012) Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells. International journal of molecular sciences, 13 (12), 15801-15812 doi:10.3390/ijms131215801.
@article{article, author = {von Einem, Bjorn and Weber, Petra and Wagner, Michael and Malnar, Martina and Ko\v{s}i\v{c}ek, Marko and He\'{c}imovi\'{c}, Silva and von Arnim, Christine and Schneckenburger, Herbert}, year = {2012}, pages = {15801-15812}, DOI = {10.3390/ijms131215801}, keywords = {FRET, cholesterol, APP, BACE1, NPC, TIRFM, neurodegeneration, Laurdan, generalized polarization}, journal = {International journal of molecular sciences}, doi = {10.3390/ijms131215801}, volume = {13}, number = {12}, issn = {1422-0067}, title = {Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells}, keyword = {FRET, cholesterol, APP, BACE1, NPC, TIRFM, neurodegeneration, Laurdan, generalized polarization} }
@article{article, author = {von Einem, Bjorn and Weber, Petra and Wagner, Michael and Malnar, Martina and Ko\v{s}i\v{c}ek, Marko and He\'{c}imovi\'{c}, Silva and von Arnim, Christine and Schneckenburger, Herbert}, year = {2012}, pages = {15801-15812}, DOI = {10.3390/ijms131215801}, keywords = {FRET, cholesterol, APP, BACE1, NPC, TIRFM, neurodegeneration, Laurdan, generalized polarization}, journal = {International journal of molecular sciences}, doi = {10.3390/ijms131215801}, volume = {13}, number = {12}, issn = {1422-0067}, title = {Cholesterol dependent energy transfer between fluorescent proteins - insights into protein proximity of APP and BACE1 in different membranes in Niemann-Pick type C disease cells}, keyword = {FRET, cholesterol, APP, BACE1, NPC, TIRFM, neurodegeneration, Laurdan, generalized polarization} }

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

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