Tiede, C, Tang, AA, Deacon, Sarah E., Mandal, U, Nettleship, JE, Owen, RL, George, SE, Harrison, DJ, Owens, RJ, Tomlinson, DC and McPherson, MJ (2014) Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications. Protein Engineering Design and Selection, 27 (5). pp. 145-155. ISSN 1741-0126,
|
PDF
- Published Version
Available under License Creative Commons Attribution. Download (699kB) | Preview |
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.
Item Type: | Article |
---|---|
Subjects: | Q Science > Q Science (General) R Medicine > R Medicine (General) R Medicine > RM Therapeutics. Pharmacology |
Schools: | Huddersfield Business School |
Depositing User: | Sarah Deacon |
Date Deposited: | 20 Oct 2016 13:44 |
Last Modified: | 28 Aug 2021 16:40 |
URI: | http://eprints.hud.ac.uk/id/eprint/29681 |
Downloads
Downloads per month over past year
Repository Staff Only: item control page
![]() |
View Item |