Structural basis of LaDR5, a novel agonistic anti-death receptor 5 (DR5) monoclonal antibody, to inhibit DR5/TRAIL complex formation.
BACKGROUND
As a member of the TNF superfamily, TRAIL could induce human tumor cell apoptosis through its cognate death receptors DR4 or DR5, which can induce formation of the death inducing signaling complex (DISC) and activation of the membrane proximal caspases (caspase-8 or caspase-10) and mitochondrial pathway. Some monoclonal antibodies against DR4 or DR5 have been reported to have anti-tumor activity.


RESULTS
In this study, we reported a novel mouse anti-human DR5 monoclonal antibody, named as LaDR5, which could compete with TRAIL to bind DR5 and induce the apoptosis of Jurkat cells in the absence of second cross-linking in vitro. Using computer-guided molecular modeling method, the 3-D structure of LaDR5 Fv fragment was constructed. According to the crystal structure of DR5, the 3-D complex structure of DR5 and LaDR5 was modeled using molecular docking method. Based on distance geometry method and intermolecular hydrogen bonding analysis, the key functional domain in DR5 was predicted and the DR5 mutants were designed. And then, three mutants of DR5 was expressed in prokaryotic system and purified by affinity chromatograph to determine the epitope of DR5 identified by LaDR5, which was consistent with the theoretical results of computer-aided analysis.


CONCLUSIONS
Our results demonstrated the specific epitope located in DR5 that plays a crucial role in antibody binding and even antineoplastic bioactivity. Meanwhile, revealed structural features of DR5 may be important to design or screen novel drugs agonist DR5.
Department of Immunology, Institute of Basic Medical Sciences, Beijing, 100850, People's Republic of China.
Chunxia
C
Qiao
Meiyun
M
Hu
Leiming
L
Guo
Ming
M
Lv
Zhou
Z
Lin
Jing
J
Geng
Xiaoling
X
Lang
Xinying
X
Li
Yan
Y
Li
Yuanfang
Y
Ma
Jiannan
J
Feng
Beifen
B
Shen
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology; D017209:Apoptosis; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D018604:Epitope Mapping; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology*; D019169:Jurkat Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D008968:Molecular Conformation; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000819:agonists* / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D007135:Immunoglobulin Variable Region; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand; D053221:TNF-Related Apoptosis-Inducing Ligand