Functional domain analysis of SOX18 transcription factor using a single-chain variable fragment-based approach
Autor: | Emma Sierecki, Stephen M. Mahler, Yann Gambin, Jeremy W. Prokop, Mathias Francois, Geoffrey W. Osborne, Daniel T. Rasicci, Martina L. Jones, Christopher B. Howard, Mehdi Moustaqil, Johannes Zuegg, Stephen Francis Goodall, Frank Fontaine, Sumukh Kumble |
---|---|
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Immunology Computational biology scFv Protein–protein interaction protein-protein interaction 03 medical and health sciences Transcription (biology) Report antibody transcriptional activation SOXF Transcription Factors Humans Immunology and Allergy Single-chain variable fragment Electrophoretic mobility shift assay Mode of action Transcription factor biology Chemistry SOX18 transcription factor In vitro 030104 developmental biology biology.protein Antibody Single-Chain Antibodies |
Zdroj: | mAbs |
ISSN: | 1942-0870 1942-0862 |
Popis: | Antibodies are routinely used to study the activity of transcription factors, using various in vitro and in vivo approaches such as electrophoretic mobility shift assay, enzyme-linked immunosorbent assay, genome-wide method analysis coupled with next generation sequencing, or mass spectrometry. More recently, a new application for antibodies has emerged as crystallisation scaffolds for difficult to crystallise proteins, such as transcription factors. Only in a few rare cases, antibodies have been used to modulate the activity of transcription factors, and there is a real gap in our knowledge on how to efficiently design antibodies to interfere with transcription. The molecular function of transcription factors is underpinned by complex networks of protein-protein interaction and in theory, setting aside intra-cellular delivery challenges, developing antibody-based approaches to modulate transcription factor activity appears a viable option. Here, we demonstrate that antibodies or an antibody single-chain variable region fragments are powerful molecular tools to unravel complex protein-DNA and protein-protein binding mechanisms. In this study, we focus on the molecular mode of action of the transcription factor SOX18, a key modulator of endothelial cell fate during development, as well as an attractive target in certain pathophysiological conditions such as solid cancer metastasis. The engineered antibody we designed inhibits SOX18 transcriptional activity, by interfering specifically with an 8-amino-acid motif in the C-terminal region directly adjacent to α-Helix 3 of SOX18 HMG domain, thereby disrupting protein-protein interaction. This new approach establishes a framework to guide the study of transcription factors interactomes using antibodies as molecular handles. |
Databáze: | OpenAIRE |
Externí odkaz: |