Selection of Ankyrin Targeting HIV-1 Matrix and Identification of Its Binding Domain

Thongkum, Weeraya and Samerjai, Kanokwan and Saoin, Somphot and Wisitponchai, Tanchanok and Hadpech, Sudarat and Lee, Vannajan Sanghiran and Lim, Theam Soon and Tayapiwatana, Chatchai (2018) Selection of Ankyrin Targeting HIV-1 Matrix and Identification of Its Binding Domain. Chiang Mai University Journal of Natural Sciences, 17 (4). pp. 339-362. ISSN 1685-1994, DOI

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Ankyrin repeat protein is a novel class of non-antibody binding protein that can be applied as an alternative antiretroviral agent. Engineered ankyrin targeting the HIV-1 matrix (MA) would be a promising agent to interfere with HIV replication, since MA plays a major role in multiple processes of the viral life cycle. In this study, MA-specific ankyrin (AnkGAGG31) was isolated from an artificial ankyrin library using a semi-automated selection process with biotinylated MA-streptavidin magnetic beads. The AnkGAGG31-recognition site on MA was determined using both indirect and competitive ELISAs with overlapping MA tri-helical fragments and pentadecapeptides. The AnkGAGG31 showed the highest binding signal to the MA-fragments covering helices 2-3-4 and peptides corresponding to helix 2 (residues 25-43), which were found as the target epitope. This finding was further analyzed by molecular modeling and docking. The rational models of AnkGAGG31-MA complex indicated that the strong binding interaction was shown on helix 2 at key residues K27MA, K30M, and K32MA. Taken together, the identification of the binding domain on the MA target improves our understanding of the AnkGAGG31-MA interaction and provides the information necessary to design innovative protein targeting of the MA protein.

Item Type: Article
Uncontrolled Keywords: Ankyrin; Binding domain; HIV-1 matrix; MA protein; Phage-displayed panning
Subjects: Q Science > Q Science (General)
Q Science > QD Chemistry
Divisions: Faculty of Science > Department of Chemistry
Depositing User: Ms. Juhaida Abd Rahim
Date Deposited: 25 Feb 2019 03:57
Last Modified: 25 Feb 2019 03:57

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