YIA Session (15 min) The 48th Lorne Conference on Protein Structure and Function 2023

Secreted dengue virus NS1 is predominantly dimeric and in complex with high-density lipoprotein (#19)

Wint Wint Phoo 1 , Bing Liang Alvin Chew 2 3 4 , An Qi Ngoh 1 , Wing Ki Kitti Chan 1 , Zheng Ser 5 , Shaio See Lim 1 , Satoru Watanabe 1 , Ming Ju Milly Choy 1 , Guek Hong Jenny Low 1 6 , Eng Eong Ooi 1 6 7 , Radoslaw Sobota 5 , Dahai Luo 2 3 4 , Subhash Vasudevan 1 8
  1. Emerging Infectious Diseases , Duke-NUS Graduate Medical School, Singapore
  2. School of Biological Sciences, Nanyang Technological University, Singapore
  3. Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore
  4. Nanyang Institute of Structural Biology, Nanyang Technological University, Singapore
  5. Functional Proteomics Laboratory, Institute of Molecular and Cell Biology, Singapore
  6. Department of Infectious Diseases, Singapore General Hospital, Singapore
  7. Saw Swee Hock School of Public Health, National University of Singapore, Singapore
  8. Institute for Glycomics, Griffith University, Gold Coast, QLD, Australia

Severe dengue infections are characterized by endothelial dysfunction associated with the secreted nonstructural protein 1 (sNS1), making it an attractive vaccine antigen and therapeutic target. The sNS1 was reported to be a hexamer with a lipid containing core based on low resolution EM data of recombinant NS1 protein. However, how the sNS1 hexamer forms and how sNS1 may cause vascular leakage remains elusive. The hexamer model is challenged by recent evidence suggesting NS1 may be associated with HDL and uses scavenger receptor B1 (SCARB1) as a cell receptor in cultured cells. To reconcile the findings of the functional forms of sNS1 and better understand its biological roles, the native form of sNS1 from cells infected with DENV are purified. We determined the cryoEM structures of sNS1 and its complex with a monoclonal antibody/Fab. The major species of sNS1 is a 1:1 complex of the NS1 dimer embedded a High-Density Lipoprotein (HDL) particle. Cross-linking MS studies confirm NS1:ApoA1 dimer formation with most ApoA1 interaction sites mapped to the NS1 wing and hydrophobic domains. These results report the molecular architecture of the secreted NS1 and provide insights on the molecular mechanisms of dengue pathogenesis. 

 

  1. Secreted dengue virus NS1 is predominantly dimeric and in complex with high-density lipoprotein Bing Liang Alvin Chew*, An Qi Ngoh*, Wint Wint Phoo*, Wing Ki Kitti Chan*, Zheng Ser, Shiao See Lim, Meng Jie Grace Weng, Satoru Watanabe, Milly M. Choy, Jenny G. Low, Eng Eong Ooi, Radoslaw M. Sobota, Subhash G. Vasudevan, Dahai Luo. *Equal contributing authors bioRxiv 2022.04.06.487425; doi: https://doi.org/10.1101/2022.04.06.487425