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Open Access Highly Accessed Research article

Structural basis of LaDR5, a novel agonistic anti-death receptor 5 (DR5) monoclonal antibody, to inhibit DR5/TRAIL complex formation

Chunxia Qiao1, Meiyun Hu12, Leiming Guo1, Ming Lv1, Zhou Lin1, Jing Geng1, Xiaoling Lang1, Xinying Li1, Yan Li1, Yuanfang Ma2, Jiannan Feng1* and Beifen Shen1*

Author Affiliations

1 Department of Immunology, Institute of Basic Medical Sciences, P.O. Box130(3), Taiping Road, Beijing, 100850, People’s Republic of China

2 Laboratory of Cellular and Molecular Immunology, Institute of Immunology, Henan University, Kaifeng, 475001, People’s Republic of China

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BMC Immunology 2012, 13:40  doi:10.1186/1471-2172-13-40

Published: 12 July 2012

Abstract

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.

Keywords:
TRAIL; Death receptor 5; Monoclonal antibody; Apoptosis; Breast cancer