Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications
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...
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| Format: | Online |
| Language: | English |
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Oxford University Press
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4000234/ |
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pubmed-4000234 |
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pubmed-40002342014-06-18 Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications Tiede, Christian Tang, Anna A. S. Deacon, Sarah E. Mandal, Upasana Nettleship, Joanne E. Owen, Robin L. George, Suja E. Harrison, David J. Owens, Raymond J. Tomlinson, Darren C. McPherson, Michael J. Original Articles 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. Oxford University Press 2014-05 2014-03-25 /pmc/articles/PMC4000234/ /pubmed/24668773 http://dx.doi.org/10.1093/protein/gzu007 Text en © The Author 2014. Published by Oxford University Press. http://creativecommons.org/licenses/by/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Tiede, Christian Tang, Anna A. S. Deacon, Sarah E. Mandal, Upasana Nettleship, Joanne E. Owen, Robin L. George, Suja E. Harrison, David J. Owens, Raymond J. Tomlinson, Darren C. McPherson, Michael J. |
| spellingShingle |
Tiede, Christian Tang, Anna A. S. Deacon, Sarah E. Mandal, Upasana Nettleship, Joanne E. Owen, Robin L. George, Suja E. Harrison, David J. Owens, Raymond J. Tomlinson, Darren C. McPherson, Michael J. Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| author_facet |
Tiede, Christian Tang, Anna A. S. Deacon, Sarah E. Mandal, Upasana Nettleship, Joanne E. Owen, Robin L. George, Suja E. Harrison, David J. Owens, Raymond J. Tomlinson, Darren C. McPherson, Michael J. |
| author_sort |
Tiede, Christian |
| title |
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| title_short |
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| title_full |
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| title_fullStr |
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| title_full_unstemmed |
Adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| title_sort |
adhiron: a stable and versatile peptide display scaffold for molecular recognition applications |
| description |
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. |
| publisher |
Oxford University Press |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4000234/ |
| _version_ |
1612082646092873728 |