{"id":5361,"date":"2026-02-25T09:30:33","date_gmt":"2026-02-25T02:30:33","guid":{"rendered":"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/?p=5361"},"modified":"2026-03-30T13:29:21","modified_gmt":"2026-03-30T06:29:21","slug":"biological-properties-of-reverse-ankyrin-engineered-for-dimer-construction-to-enhance-hiv-1-capsid-interaction","status":"publish","type":"post","link":"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/biological-properties-of-reverse-ankyrin-engineered-for-dimer-construction-to-enhance-hiv-1-capsid-interaction\/","title":{"rendered":"Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction"},"content":{"rendered":"<p>On-Anong Juntit, Umpa Yasamut, Supachai Sakkhachornphop, Koollawat Chupradit, Weeraya Thongkum, <strong>Chatchawan Srisawat<\/strong>, Tawan Chokepaichitkool, Prachya Kongtawelert, Chatchai Tayapiwatana<\/p>\n<h2 class=\"title\">Abstract<\/h2>\n<div id=\"eng-abstract\" class=\"abstract-content selected\">\n<div class=\"title\">\n<p>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.<\/p>\n<p>Objective: This study aimed to improve the avidity of Ank1D4 by generating two Ank1D4 dimers: (Ank1D4NC-NC) and its inverted form (Ank1D4NC-CN), with each domain connected by a flexible (G4S)4 linker peptide.<\/p>\n<p>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. Ank1D4NC-NC and Ank1D4NC-CN were evaluated using bio-layer interferometry (BLI), compared to monomeric Ank1D4.<\/p>\n<p>Results: Similar binding surfaces were observed in both dimers which was comparable with monomeric Ank1D4. The interaction of Ank1D4NC-CN with CAp24 was significantly greater than that of Ank1D4NC-NC and Ank1D4 by capture ELISA. Ank1D4NC-CN also exhibited bifunctionality using a sandwich ELISA. The KD of Ank1D4NC-CN, Ank1D4NC-NC and monomeric Ank1D4 was 3.5 nM, 53.7 nM, and 126.2 nM, respectively using bio-layer interferometry analysis.<\/p>\n<p>Conclusions: This study provides a strategy for increasing Ank1D4 avidity through the construction of novel inverted dimers with a flexible linker. Ank1D4NC-CN may provide an alternative treatment strategy for inhibiting HIV-1 replication.<\/p>\n<p><a href=\"https:\/\/pubmed.ncbi.nlm.nih.gov\/35598193\/\">https:\/\/pubmed.ncbi.nlm.nih.gov\/35598193\/<\/a><\/p>\n<\/div>\n<\/div>\n","protected":false},"excerpt":{"rendered":"<p>On-Anong Juntit, Umpa Yasamut, Supachai Sakkhachornphop, Koollawat Chupradit, Weeraya Thongkum, Chatchawan Srisawat, Tawan Chokepaichitkool, Prachya Kongtawelert, Chatchai Tayapiwatana 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 [&#8230;]\n","protected":false},"author":6,"featured_media":0,"comment_status":"closed","ping_status":"closed","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[18],"tags":[],"class_list":["post-5361","post","type-post","status-publish","format-standard","hentry","category-research-innovation-en"],"yoast_head":"<!-- This site is optimized with the Yoast SEO plugin v27.6 - https:\/\/yoast.com\/product\/yoast-seo-wordpress\/ -->\n<title>Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction - \u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/biological-properties-of-reverse-ankyrin-engineered-for-dimer-construction-to-enhance-hiv-1-capsid-interaction\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Biological properties of reverse ankyrin engineered for dimer construction to enhance HIV-1 capsid interaction - \u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25\" \/>\n<meta property=\"og:description\" content=\"On-Anong Juntit, Umpa Yasamut, Supachai Sakkhachornphop, Koollawat Chupradit, Weeraya Thongkum, Chatchawan Srisawat, Tawan Chokepaichitkool, Prachya Kongtawelert, Chatchai Tayapiwatana 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. 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We previously generated a monomeric ankyrin repeat protein [...]","og_url":"https:\/\/www2.si.mahidol.ac.th\/department\/biochemistry\/en\/biological-properties-of-reverse-ankyrin-engineered-for-dimer-construction-to-enhance-hiv-1-capsid-interaction\/","og_site_name":"\u0e20\u0e32\u0e04\u0e27\u0e34\u0e0a\u0e32\u0e0a\u0e35\u0e27\u0e40\u0e04\u0e21\u0e35 \u0e04\u0e13\u0e30\u0e41\u0e1e\u0e17\u0e22\u0e28\u0e32\u0e2a\u0e15\u0e23\u0e4c\u0e28\u0e34\u0e23\u0e34\u0e23\u0e32\u0e0a\u0e1e\u0e22\u0e32\u0e1a\u0e32\u0e25 \u0e21\u0e2b\u0e32\u0e27\u0e34\u0e17\u0e22\u0e32\u0e25\u0e31\u0e22\u0e21\u0e2b\u0e34\u0e14\u0e25","article_publisher":"https:\/\/www.facebook.com\/SIBBprogram\/","article_published_time":"2026-02-25T02:30:33+00:00","article_modified_time":"2026-03-30T06:29:21+00:00","author":"Poorintra Sakuna","twitter_card":"summary_large_image","twitter_misc":{"Written by":"Poorintra Sakuna","Est. reading time":"2 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