Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction
Issued Date
2025-09-01
Resource Type
ISSN
0125877X
eISSN
22288694
Scopus ID
2-s2.0-105020826365
Pubmed ID
35598193
Journal Title
Asian Pacific Journal of Allergy and Immunology
Volume
43
Issue
3
Start Page
737
End Page
744
Rights Holder(s)
SCOPUS
Bibliographic Citation
Asian Pacific Journal of Allergy and Immunology Vol.43 No.3 (2025) , 737-744
Suggested Citation
Juntit O.A., Yasamut U., Sakkhachornphop S., Chupradit K., Thongkum W., Srisawat C., Chokepaichitkool T., Kongtawelert P., Tayapiwatana C. Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction. Asian Pacific Journal of Allergy and Immunology Vol.43 No.3 (2025) , 737-744. 744. doi:10.12932/ap-210122-1310 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/112990
Title
Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction
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Abstract
Background: Assembly and budding in the late-stage of human immunodeficiency virus type 1 (HIV-1) production rely on Gag protein polymerization at the inner leaflet of the plasma membrane. We previously generated a monomeric ankyrin repeat protein (Ank1D4) that specifically interacts with capsid protein (CAp24) of HIV-1, however this protein had modest binding affinity. Objective: This study aimed to improve the avidity of Ank1D4 by generating two Ank1D4 dimers: (Ank1D4<inf>NC-NC</inf>) and its inverted form (Ank1D4<inf>NC-CN</inf>), with each domain connected by a flexible (G<inf>4</inf> S)<inf>4</inf> linker peptide. Methods: Binding properties of monomeric and dimeric Ank1D4 was performed by capture enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA was used to examine bifunctional module of dimeric Ank1D4. Ank1D4<inf>NC-NC</inf> and Ank1D4<inf>NC-CN</inf> were evaluated using bio-layer interferometry (BLI), compared to monomeric Ank1D4. Results: Similar binding surfaces were observed in both dimers which was comparable with monomeric Ank1D4. The interaction of Ank1D4<inf>NC-CN</inf> with CAp24 was significantly greater than that of Ank1D4<inf>NC-NC</inf> and Ank1D4 by capture ELISA. Ank1D4<inf>NC-CN</inf> also exhibited bifunctionality using a sandwich ELISA. The KD of Ank1D4<inf>NC-CN</inf>, Ank1D4<inf>NC-NC</inf> and monomeric Ank1D4 was 3.5 nM, 53.7 nM, and 126.2 nM, respectively using bio-layer interferometry analysis. Conclusions: This study provides a strategy for increasing Ank1D4 avidity through the construction of novel inverted dimers with a flexible linker. Ank1D4<inf>NC-CN</inf> may provide an alternative treatment strategy for inhibiting HIV-1 replication.