We analyzed a novel bifunctional fusion protein, CD40ed-CD95Led, consisting amino-terminally of the extracellular domain of CD40 and carboxy-terminally of the extracellular domain of CD95L. On cells lacking CD40L, this fusion protein is poorly active with respect to CD95 activation median effective dose (ED50)\textgreater1 microg/ml, but it stimulates CD95 signaling with high efficiency upon binding to membrane-expressed CD40L (ED50\textless1 ng/ml). Thus, cell surface immobilization mediated by the CD40 part of the molecule unmasks the high-latent, CD95-stimulating capacity of the otherwise poorly active CD95L fusion protein. Moreover, interaction of the CD40 part of CD40ed-CD95Led with CD40L prevents the activation of cellular CD40. The CD40ed-CD95Led fusion protein therefore simultaneously blocks antiapoptotic CD40 activation and induces CD95-mediated apoptosis. Indeed, T47D cells displaying an antiapoptotic autocrine CD40-CD40L signaling loop were significantly more sensitive toward CD40ed-CD95Led than toward soluble CD95L artificially activated by crosslinking. Fusion proteins of RANK and CD95L (RANKed-CD95Led) and CD40 and tumor necrosis factor-related apoptosis inducing ligand (TRAIL) (CD40ed-TRAILed), with domain architectures similar to CD40ed-Cd95Led, displayed RANKL-dependent CD95 and CD40L-dependent TRAILR2 activation, respectively, indicating the principle feasibility of this fusion protein design.
%0 Journal Article
%1 Assohou-Luty2006
%A Assohou-Luty, Constance
%A Gerspach, Jeanette
%A Siegmund, Daniela
%A Müller, Nicole
%A Huard, Bertrand
%A Tiegs, Gisa
%A Pfizenmaier, Klaus
%A Wajant, Harald
%D 2006
%J Journal of Molecular Medicine
%K 2006 izi pfizenmaier
%N 9
%P 785--797
%R 10.1007/s00109-006-0073-1
%T A CD40-CD95L fusion protein interferes with CD40L-induced prosurvival signaling and allows membrane CD40L-restricted activation of CD95
%U https://www.ncbi.nlm.nih.gov/pubmed/16924474
%V 84
%X We analyzed a novel bifunctional fusion protein, CD40ed-CD95Led, consisting amino-terminally of the extracellular domain of CD40 and carboxy-terminally of the extracellular domain of CD95L. On cells lacking CD40L, this fusion protein is poorly active with respect to CD95 activation median effective dose (ED50)\textgreater1 microg/ml, but it stimulates CD95 signaling with high efficiency upon binding to membrane-expressed CD40L (ED50\textless1 ng/ml). Thus, cell surface immobilization mediated by the CD40 part of the molecule unmasks the high-latent, CD95-stimulating capacity of the otherwise poorly active CD95L fusion protein. Moreover, interaction of the CD40 part of CD40ed-CD95Led with CD40L prevents the activation of cellular CD40. The CD40ed-CD95Led fusion protein therefore simultaneously blocks antiapoptotic CD40 activation and induces CD95-mediated apoptosis. Indeed, T47D cells displaying an antiapoptotic autocrine CD40-CD40L signaling loop were significantly more sensitive toward CD40ed-CD95Led than toward soluble CD95L artificially activated by crosslinking. Fusion proteins of RANK and CD95L (RANKed-CD95Led) and CD40 and tumor necrosis factor-related apoptosis inducing ligand (TRAIL) (CD40ed-TRAILed), with domain architectures similar to CD40ed-Cd95Led, displayed RANKL-dependent CD95 and CD40L-dependent TRAILR2 activation, respectively, indicating the principle feasibility of this fusion protein design.
%7 2006/08/23
%@ 0946-2716 (Print)$\backslash$r0946-2716 (Linking)
@article{Assohou-Luty2006,
abstract = {We analyzed a novel bifunctional fusion protein, CD40ed-CD95Led, consisting amino-terminally of the extracellular domain of CD40 and carboxy-terminally of the extracellular domain of CD95L. On cells lacking CD40L, this fusion protein is poorly active with respect to CD95 activation [median effective dose (ED50){\textgreater}1 microg/ml], but it stimulates CD95 signaling with high efficiency upon binding to membrane-expressed CD40L (ED50{\textless}1 ng/ml). Thus, cell surface immobilization mediated by the CD40 part of the molecule unmasks the high-latent, CD95-stimulating capacity of the otherwise poorly active CD95L fusion protein. Moreover, interaction of the CD40 part of CD40ed-CD95Led with CD40L prevents the activation of cellular CD40. The CD40ed-CD95Led fusion protein therefore simultaneously blocks antiapoptotic CD40 activation and induces CD95-mediated apoptosis. Indeed, T47D cells displaying an antiapoptotic autocrine CD40-CD40L signaling loop were significantly more sensitive toward CD40ed-CD95Led than toward soluble CD95L artificially activated by crosslinking. Fusion proteins of RANK and CD95L (RANKed-CD95Led) and CD40 and tumor necrosis factor-related apoptosis inducing ligand (TRAIL) (CD40ed-TRAILed), with domain architectures similar to CD40ed-Cd95Led, displayed RANKL-dependent CD95 and CD40L-dependent TRAILR2 activation, respectively, indicating the principle feasibility of this fusion protein design.},
added-at = {2018-02-01T15:53:05.000+0100},
author = {Assohou-Luty, Constance and Gerspach, Jeanette and Siegmund, Daniela and M{\"{u}}ller, Nicole and Huard, Bertrand and Tiegs, Gisa and Pfizenmaier, Klaus and Wajant, Harald},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/2ea7ad1b4d52c672f36e360d387760a0e/cristiano},
doi = {10.1007/s00109-006-0073-1},
edition = {2006/08/23},
interhash = {b7520997ca1a6529152830ba3bae780a},
intrahash = {ea7ad1b4d52c672f36e360d387760a0e},
isbn = {0946-2716 (Print)$\backslash$r0946-2716 (Linking)},
issn = {09462716},
journal = {Journal of Molecular Medicine},
keywords = {2006 izi pfizenmaier},
number = 9,
pages = {785--797},
pmid = {16924474},
timestamp = {2018-07-25T12:33:08.000+0200},
title = {{A CD40-CD95L fusion protein interferes with CD40L-induced prosurvival signaling and allows membrane CD40L-restricted activation of CD95}},
url = {https://www.ncbi.nlm.nih.gov/pubmed/16924474},
volume = 84,
year = 2006
}
