Majkut, J;
Sgobba, M;
Holohan, C;
Crawford, N;
Logan, AE;
Kerr, E;
Higgins, CA;
... Longley, DB; + view all
(2014)
Differential affinity of FLIP and procaspase 8 for FADD's DED binding surfaces regulates DISC assembly.
Nature Communications
, 5
, Article 3350. 10.1038/ncomms4350.
Preview |
Text
Majkut_differential_affinity_of_FLIP_and_procaspase.pdf Download (1MB) | Preview |
Abstract
Death receptor activation triggers recruitment of FADD, which via its death effector domain (DED) engages the DEDs of procaspase 8 and its inhibitor FLIP to form death-inducing signalling complexes (DISCs). The DEDs of FADD, FLIP and procaspase 8 interact with one another using two binding surfaces defined by α1/α4 and α2/α5 helices, respectively. Here we report that FLIP has preferential affinity for the α1/α4 surface of FADD, whereas procaspase 8 has preferential affinity for FADD's α2/α5 surface. These relative affinities contribute to FLIP being recruited to the DISC at comparable levels to procaspase 8 despite lower cellular expression. Additional studies, including assessment of DISC stoichiometry and functional assays, suggest that following death receptor recruitment, the FADD DED preferentially engages FLIP using its α1/α4 surface and procaspase 8 using its α2/α5 surface; these tripartite intermediates then interact via the α1/α4 surface of FLIP DED1 and the α2/α5 surface of procaspase 8 DED2.
| Type: | Article |
|---|---|
| Title: | Differential affinity of FLIP and procaspase 8 for FADD's DED binding surfaces regulates DISC assembly. |
| Location: | England |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1038/ncomms4350 |
| Publisher version: | http://dx.doi.org/10.1038/ncomms4350 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Blotting, Western, CASP8 and FADD-Like Apoptosis Regulating Protein, Caspase 8, Chromatography, Gel, Fas-Associated Death Domain Protein, HCT116 Cells, Humans, Immunoprecipitation, Protein Binding |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > UCL School of Pharmacy > Pharma and Bio Chemistry |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1487234 |
Archive Staff Only
 |
View Item |
