Nat Struct Mol Biol. 2010 Nov;17(11):1324-9. Epub 2010 Oct 10.

The Fas-FADD death domain complex structure reveals the basis of DISC assembly and disease mutations.External

Wang, L., Yang, J. K., Kabaleeswaran, V., Rice, A. J., Cruz, A. C., Park, A. Y., Yin, Q., Damko, E., Jang, S. B., Raunser, S., Robinson, C. V., Siegel, R. M., Walz, T., Wu, H.,
--- - Department of Biochemistry, Weill Cornell Medical College, New York, New York, USA.
The death-inducing signaling complex (DISC) formed by the death receptor Fas, the adaptor protein FADD and caspase-8 mediates the extrinsic apoptotic program. Mutations in Fas that disrupt the DISC cause autoimmune lymphoproliferative syndrome (ALPS). Here we show that the Fas-FADD death domain (DD) complex forms an asymmetric oligomeric structure composed of 5-7 Fas DD and 5 FADD DD, whose interfaces harbor ALPS-associated mutations. Structure-based mutations disrupt the Fas-FADD interaction in vitro and in living cells; the severity of a mutation correlates with the number of occurrences of a particular interaction in the structure. The highly oligomeric structure explains the requirement for hexameric or membrane-bound FasL in Fas signaling. It also predicts strong dominant negative effects from Fas mutations, which are confirmed by signaling assays. The structure optimally positions the FADD death effector domain (DED) to interact with the caspase-8 DED for caspase recruitment and higher-order aggregation.
PMID: 20935634External
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vitro interaction
  DD1 DD2 Reference
Family DD1 DD2 Method Species Region Expression Species Region Expression
DD FADD Link Fas Affinity purification-mass spectrometry Human 93-184 E.coli Mouse 210-310 E.coli 20935634
DD FADD Link Fas Affinity purification-mass spectrometry Human 93-191 E.coli Human 218-318 E.coli 20935634
DD FADD Link Fas His fusion protein pull-down Not specified Not specified E.coli Not specified Not specified E.coli 20935634
DD FADD Link Fas In vitro purification protein assembly(Size-exclusion chromatography) Human 93-184 E.coli Human 210-310 E.coli 20935634
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vivo interaction
  Endogenous
expression
Overexpression DD1 DD2 Reference
Family DD1 DD2 Method Species Region Species Region
DD FADD Link Fas Fluorescence resonance energy transfer HEK293 Human Not specified Human Full length 20935634
(Link: click this icon to show interactions only between the two corresponding DDs)
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
Characterization
  Stoichiometry Affinity Protein-Protein interface Reference
Family DD1 DD2 Method Mutation Complex structure
DD FADD Link Fas Mass spectrometry, MALS 5:5, 5:6, 5:7 20935634
DD FADD Link Fas Fluorescence resonance energy transfer D260V, D260Y, A257D, E261K, T270K, E272K, Q283K (Fas) R117E, D123R, R135E, R172E (FADD) 20935634
DD FADD Link Fas His fusion protein pull-down E261K, E272K, T270K, R250E, Q283K, K287D, R250Q, D260Y, A257D, E272K, T270K (fas) 20935634
DD FADD Link Fas Structure 5:5 3OQ9 20935634
(Link: click this icon to show interactions only between the two corresponding DDs)