Publication

Article Metrics

Citations


Online attention

Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications

DOI: 10.1093/protein/gzu007 DOI Help
PMID: 24668773 PMID Help

Authors: C. Tiede (University of Leeds) , A. A. S. Tang (University of Leeds) , S. E. Deacon (University of Leeds) , U. Mandal (University of Leeds) , J. E. Nettleship (University of Oxford) , Robin Owen (Diamond Light Source) , S. E. George (University of Leeds) , D. J. Harrison (University of Leeds) , Ray Owens (University of Oxford) , D. C. Tomlinson (University of Leeds) , M. J. Mcpherson (University of Leeds)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Protein Engineering Design And Selection , VOL 27 (5) , PAGES 145 - 155

State: Published (Approved)
Published: April 2014
Diamond Proposal Number(s): 5968

Open Access Open Access

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 × 1010 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

Subject Areas: Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography

Discipline Tags:



Technical Tags: