TY - JOUR
T1 - The C-terminal region of tumor necrosis factor like weak inducer of apoptosis is required for interaction with advanced glycation end products
AU - Watanabe, Masahiro
AU - Toyomura, Takao
AU - Wake, Hidenori
AU - Liu, Keyue
AU - Teshigawara, Kiyoshi
AU - Takahashi, Hideo
AU - Nishibori, Masahiro
AU - Mori, Shuji
N1 - Funding Information:
This work was supported by MEXT/JSPS KAKENHI grant numbers 18K06807 and 18K14969, the Wesco Scientific Promotion Foundation, the Ryobi Teien Memory Foundation, and the Shujitsu University Grant-in-Aid for Education, Research and Publication.
Publisher Copyright:
© 2018 International Union of Biochemistry and Molecular Biology, Inc.
PY - 2019/3/1
Y1 - 2019/3/1
N2 - Previously, we found that endogenously produced pro-inflammatory molecules, advanced glycation end products (AGEs), interact with tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and attenuate its immunomodulatory function. In the present study, to elucidate the mechanism by which AGEs attenuate TWEAK function, we searched for regions responsible for TWEAK–AGE interaction using TWEAK deletion mutants. Pull-down assays with the TWEAK mutants and AGEs revealed that the C-terminal half of TWEAK, which is the region essential for receptor stimulation, was required for this interaction. On the other hand, the N-terminal deletion mutants did not exhibit a significant decrease in AGE binding. Moreover, a moderate decrease in the AGE binding by double-deletion in quartered C-terminal half regions and a substantial decrease by triple-deletion in this region were observed. In addition, full-length TWEAK stimulated IL-8 gene expression in endothelial EA.hy.926 cells, whereas the triple-deletion mutant lost much of this activity, suggesting that the TWEAK–AGE interaction sites overlap with the region needed to exert normal function of TWEAK. Our present findings may help to elucidate the pathophysiological roles of the TWEAK–AGE interaction for prevention and treatment of AGE-related inflammatory diseases.
AB - Previously, we found that endogenously produced pro-inflammatory molecules, advanced glycation end products (AGEs), interact with tumor necrosis factor-like weak inducer of apoptosis (TWEAK), and attenuate its immunomodulatory function. In the present study, to elucidate the mechanism by which AGEs attenuate TWEAK function, we searched for regions responsible for TWEAK–AGE interaction using TWEAK deletion mutants. Pull-down assays with the TWEAK mutants and AGEs revealed that the C-terminal half of TWEAK, which is the region essential for receptor stimulation, was required for this interaction. On the other hand, the N-terminal deletion mutants did not exhibit a significant decrease in AGE binding. Moreover, a moderate decrease in the AGE binding by double-deletion in quartered C-terminal half regions and a substantial decrease by triple-deletion in this region were observed. In addition, full-length TWEAK stimulated IL-8 gene expression in endothelial EA.hy.926 cells, whereas the triple-deletion mutant lost much of this activity, suggesting that the TWEAK–AGE interaction sites overlap with the region needed to exert normal function of TWEAK. Our present findings may help to elucidate the pathophysiological roles of the TWEAK–AGE interaction for prevention and treatment of AGE-related inflammatory diseases.
KW - advanced glycation end products
KW - binding
KW - cytokine
KW - deletion mutant
KW - tumor necrosis factor like weak inducer of apoptosis
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U2 - 10.1002/bab.1706
DO - 10.1002/bab.1706
M3 - Article
C2 - 30403295
AN - SCOPUS:85058851235
VL - 66
SP - 254
EP - 260
JO - Journal of Applied Biochemistry
JF - Journal of Applied Biochemistry
SN - 0885-4513
IS - 2
ER -