Caspase-8 Binding to Cardiolipin in Giant Unilamellar Vesicles Provides a Functional Docking Platform for Bid

Olivier Jalmar, Liberty Franc¸ois-Moutal, Ana-Jesus García-Sáez, Mark Perry, Thierry Granjon, Francois Gonzalvez, Eyal Gottlieb, Jesus Ayala-Sanmartin, Beate Maria Klösgen-Buchkremer, Petra Schwille, Patrice X. Petit

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

308 Downloads (Pure)

Abstrakt

Caspase-8 is involved in death receptor-mediated apoptosis in type II cells, the proapoptotic programme of which is triggered by truncated Bid. Indeed, caspase-8 and Bid are the known intermediates of this signalling pathway. Cardiolipin has been shown to provide an anchor and an essential activating platform for caspase-8 at the mitochondrial membrane surface. Destabilisation of this platform alters receptor-mediated apoptosis in diseases such as Barth Syndrome, which is characterised by the presence of immature cardiolipin which does not allow caspase-8 binding. We used a simplified in vitro system that mimics contact sites and/or cardiolipin-enriched microdomains at the outer mitochondrial surface in which the platform consisting of caspase-8, Bid and cardiolipin was reconstituted in giant unilamellar vesicles. We analysed these vesicles by flow cytometry and confirm previous results that demonstrate the requirement for intact mature cardiolipin for caspase-8 activation and Bid binding and cleavage. We also used confocal microscopy to visualise the rupture of the vesicles and their revesiculation at smaller sizes due to alteration of the curvature following caspase-8 and Bid binding. Biophysical approaches, including Laurdan fluorescence and rupture/tension measurements, were used to determine the ability of these three components (cardiolipin, caspase-8 and Bid) to fulfil the minimal requirements for the formation and function of the platform at the mitochondrial membrane. Our results shed light on the active functional role of cardiolipin, bridging the gap between death receptors and mitochondria.

OriginalsprogEngelsk
Artikelnummere55250
TidsskriftPLOS ONE
Vol/bind8
Udgave nummer2
Sider (fra-til)1-12
Antal sider12
ISSN1932-6203
DOI
StatusUdgivet - 13. feb. 2013

Fingeraftryk

Dyk ned i forskningsemnerne om 'Caspase-8 Binding to Cardiolipin in Giant Unilamellar Vesicles Provides a Functional Docking Platform for Bid'. Sammen danner de et unikt fingeraftryk.

Citationsformater