Eur J Biochem. 2001 Mar;268(5):1382-91.

Structure-activity relationship of the p55 TNF receptor death domain and its lymphoproliferation mutants.External

De Wilde, G., Murray-Rust, J., Boone, E., Olerenshaw, D., McDonald, N. Q., Ibanez, C., Haegeman, G., Wollmer, A., Federwisch, M.,
--- - Department of Molecular Biology, University of Gent-VIB, Belgium.
Upon stimulation with tumor necrosis factor (TNF), the TNF receptor (TNFR55) mediates a multitude of effects both in normal and in tumor cells. Clustering of the intracellular domain of the receptor, the so-called death domain (DD), is responsible for both the initiation of cell killing and the activation of gene expression. To characterize this domain further, TNFR55 DD was expressed and purified as a thioredoxin fusion protein in Escherichia coli. Circular dichroism, steady-state and time-resolved fluorescence spectroscopy were used to compare TNFR55 DD with DDs of the Fas antigen (Fas), the Fas-associating protein with DD (FADD) and p75 nerve growth factor receptor, for which the 3-dimensional structure are already known. The structural information derived from the measurements strongly suggests that TNFR55 DD adopts a similar fold in solution. This prompted a homology modeling of the TNFR DD 3-D structure using FADD as a template. In vivo studies revealed a difference between the two lymphoproliferation (lpr) mutations. Biophysical techniques were used to analyze the effect of changing Leu351 to Ala and Leu351 to Asn on the global structure and its impact on the overall stability of TNFR55 DD. The results obtained from these experiments in combination with the modeled structure offer an explanation for the in vivo observed difference.
PMID: 11231290External
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 TNFR1 Link TNFR1 GST fusion protein pull-down Not specified 326-426 E.coli Not specified 214-455 E.coli 11231290
DD TNFR1 Link TRADD GST fusion protein pull-down Not specified 326-426 E.coli Not specified Not specified E.coli 11231290
(Link: click this icon to show interactions only between the two corresponding DDs)