4N6T

Adhiron: a stable and versatile peptide display scaffold - full length adhiron

4N6T の概要

• エントリーDOI: 10.2210/pdb4n6t/pdb
• 関連するPDBエントリー: 4N6U
• 分子名称: Adhiron (2 entities in total)
• 機能のキーワード: protein scaffold, consensus protein, de novo protein
• 由来する生物種: ARTIFICIAL GENE
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 9286.67

構造登録者

Mcpherson, M.,Tomlinson, D.,Owen, R.L.,Nettleship, J.E.,Owens, R.J. (登録日: 2013-10-14, 公開日: 2014-04-09, 最終更新日: 2024-02-28)

主引用文献

Tiede, C.,Tang, A.A.,Deacon, S.E.,Mandal, U.,Nettleship, J.E.,Owen, R.L.,George, S.E.,Harrison, D.J.,Owens, R.J.,Tomlinson, D.C.,McPherson, M.J.
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications.
Protein Eng.Des.Sel., 27:145-155, 2014

PubMed Abstract: We have designed a novel non-antibody scaffold protein, termed Adhiron, based on a phytocystatin consensus sequence. The Adhiron scaffold shows high thermal stability (Tm ca. 101°C), and is expressed well in Escherichia coli. We have determined the X-ray crystal structure of the Adhiron scaffold to 1.75 Å resolution revealing a compact cystatin-like fold. We have constructed a phage-display library in this scaffold by insertion of two variable peptide regions. The library is of high quality and complexity comprising 1.3 × 10(10) clones. To demonstrate library efficacy, we screened against the yeast Small Ubiquitin-like Modifier (SUMO). In selected clones, variable region 1 often contained sequences homologous to the known SUMO interactive motif (V/I-X-V/I-V/I). Four Adhirons were further characterised and displayed low nanomolar affinities and high specificity for yeast SUMO with essentially no cross-reactivity to human SUMO protein isoforms. We have identified binders against >100 target molecules to date including as examples, a fibroblast growth factor (FGF1), platelet endothelial cell adhesion molecule (PECAM-1; CD31), the SH2 domain Grb2 and a 12-aa peptide. Adhirons are highly stable and well expressed allowing highly specific binding reagents to be selected for use in molecular recognition applications.

PubMed: 24668773
DOI: 10.1093/protein/gzu007

実験手法

X-RAY DIFFRACTION (1.75 Å)