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Cognate peptide-receptor ligand mapping by directed phage display
BACKGROUND: A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relevant ligands. METHODS: A gene fragment library from infl...
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Formato: | Texto |
Lenguaje: | English |
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BioMed Central
2005
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1183247/ https://www.ncbi.nlm.nih.gov/pubmed/15963231 http://dx.doi.org/10.1186/1477-5956-3-7 |
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author | Stratmann, Thomas Kang, Angray S |
author_facet | Stratmann, Thomas Kang, Angray S |
author_sort | Stratmann, Thomas |
collection | PubMed |
description | BACKGROUND: A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relevant ligands. METHODS: A gene fragment library from influenza coat protein haemagglutinin (HA) gene was constructed by treating HA cDNA with DNAse I to create 50 – 100 bp fragments. These fragments were cloned into plasmid pORFES IV and in-frame inserts were selected. These in-frame fragment inserts were subsequently cloned into a filamentous phage display vector JC-M13-88 for surface display as fusions to a synthetic copy of gene VIII. Two well characterized antibodies, mAb 12CA5 and pAb 07431, directed against distinct known regions of HA were used to pan the library. RESULTS: Two linear epitopes, HA peptide 112 – 126 and 162–173, recognized by mAb 12CA5 and pAb 07431, respectively, were identified as the cognate epitopes. CONCLUSION: This approach is a useful alternative to conventional methods such as screening of overlapping synthetic peptide libraries or gene fragment expression libraries when searching for precise peptide protein interactions, and may be applied to functional proteomics. |
format | Text |
id | pubmed-1183247 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2005 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-11832472005-08-06 Cognate peptide-receptor ligand mapping by directed phage display Stratmann, Thomas Kang, Angray S Proteome Sci Methodology BACKGROUND: A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relevant ligands. METHODS: A gene fragment library from influenza coat protein haemagglutinin (HA) gene was constructed by treating HA cDNA with DNAse I to create 50 – 100 bp fragments. These fragments were cloned into plasmid pORFES IV and in-frame inserts were selected. These in-frame fragment inserts were subsequently cloned into a filamentous phage display vector JC-M13-88 for surface display as fusions to a synthetic copy of gene VIII. Two well characterized antibodies, mAb 12CA5 and pAb 07431, directed against distinct known regions of HA were used to pan the library. RESULTS: Two linear epitopes, HA peptide 112 – 126 and 162–173, recognized by mAb 12CA5 and pAb 07431, respectively, were identified as the cognate epitopes. CONCLUSION: This approach is a useful alternative to conventional methods such as screening of overlapping synthetic peptide libraries or gene fragment expression libraries when searching for precise peptide protein interactions, and may be applied to functional proteomics. BioMed Central 2005-06-17 /pmc/articles/PMC1183247/ /pubmed/15963231 http://dx.doi.org/10.1186/1477-5956-3-7 Text en Copyright © 2005 Stratmann and Kang; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Stratmann, Thomas Kang, Angray S Cognate peptide-receptor ligand mapping by directed phage display |
title | Cognate peptide-receptor ligand mapping by directed phage display |
title_full | Cognate peptide-receptor ligand mapping by directed phage display |
title_fullStr | Cognate peptide-receptor ligand mapping by directed phage display |
title_full_unstemmed | Cognate peptide-receptor ligand mapping by directed phage display |
title_short | Cognate peptide-receptor ligand mapping by directed phage display |
title_sort | cognate peptide-receptor ligand mapping by directed phage display |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1183247/ https://www.ncbi.nlm.nih.gov/pubmed/15963231 http://dx.doi.org/10.1186/1477-5956-3-7 |
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