5WYM

Crystal structure of an anti-connexin26 scFv

5WYM の概要

• エントリーDOI: 10.2210/pdb5wym/pdb
• 分子名称: anti-connexin26 scFv,Ig heavy chain,Linker,anti-connexin26 scFv,Ig light chain (2 entities in total)
• 機能のキーワード: antibody, scfv, connexin26, immune system
• 由来する生物種: Homo sapiens (Human); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 50928.68

構造登録者

Li, S.,Xu, L. (登録日: 2017-01-13, 公開日: 2018-01-24, 最終更新日: 2024-11-06)

主引用文献

Xu, L.,Carrer, A.,Zonta, F.,Qu, Z.,Ma, P.,Li, S.,Ceriani, F.,Buratto, D.,Crispino, G.,Zorzi, V.,Ziraldo, G.,Bruno, F.,Nardin, C.,Peres, C.,Mazzarda, F.,Salvatore, A.M.,Raspa, M.,Scavizzi, F.,Chu, Y.,Xie, S.,Yang, X.,Liao, J.,Liu, X.,Wang, W.,Wang, S.,Yang, G.,Lerner, R.A.,Mammano, F.
Design and Characterization of a Human Monoclonal Antibody that Modulates Mutant Connexin 26 Hemichannels Implicated in Deafness and Skin Disorders
Front Mol Neurosci, 10:298-298, 2017

PubMed Abstract: Mutations leading to changes in properties, regulation, or expression of connexin-made channels have been implicated in 28 distinct human hereditary diseases. Eight of these result from variants of connexin 26 (Cx26), a protein critically involved in cell-cell signaling in the inner ear and skin. Lack of non-toxic drugs with defined mechanisms of action poses a serious obstacle to therapeutic interventions for diseases caused by mutant connexins. In particular, molecules that specifically modulate connexin hemichannel function without affecting gap junction channels are considered of primary importance for the study of connexin hemichannel role in physiological as well as pathological conditions. Monoclonal antibodies developed in the last three decades have become the most important class of therapeutic biologicals. Recombinant methods permit rapid selection and improvement of monoclonal antibodies from libraries with large diversity. By screening a combinatorial library of human single-chain fragment variable (scFv) antibodies expressed in phage, we identified a candidate that binds an extracellular epitope of Cx26. We characterized antibody action using a variety of biochemical and biophysical assays in HeLa cells, organotypic cultures of mouse cochlea and human keratinocyte-derived cells. We determined that the antibody is a remarkably efficient, non-toxic, and completely reversible inhibitor of hemichannels formed by connexin 26 and does not affect direct cell-cell communication via gap junction channels. Importantly, we also demonstrate that the antibody efficiently inhibits hyperative mutant Cx26 hemichannels implicated in autosomal dominant non-syndromic hearing impairment accompanied by keratitis and hystrix-like ichthyosis-deafness (KID/HID) syndrome. We solved the crystal structure of the antibody, identified residues that are critical for binding and used molecular dynamics to uncover its mechanism of action. Although further studies will be necessary to validate the effect of the antibody , the methodology described here can be extended to select antibodies against hemichannels composed by other connexin isoforms and, consequently, to target other pathologies associated with hyperactive hemichannels. Our study highlights the potential of this approach and identifies connexins as therapeutic targets addressable by screening phage display libraries expressing human randomized antibodies.

PubMed: 29018324
DOI: 10.3389/fnmol.2017.00298

実験手法

X-RAY DIFFRACTION (2.65 Å)