Decoding a PNA encoded peptide library by PCR: the discovery of new cell surface receptor ligands.
The ability to screen and identify new ligands for cell surface receptors has been a long-standing goal as it might allow targeting of pharmaceutically relevant receptors, such as integrins or G protein coupled receptors. Here, we present a method to amplify hits from a library of PNA-tagged peptides. To this end, human cells, overexpressing either integrins or the CCR6 receptor, were treated with a 10,000 member PNA-encoded peptide library. Extraction of the PNA tags from the surface of the cells was followed by a PNA-tag to DNA translation and amplification enabling decoding of the tags via microarray hybridization. This approach to ligand discovery facilitates screening for differences in surface-receptor ligands and/or receptor expression between different cell types, and opens up a practical approach to PNA-tag amplification.
School of Chemistry, University of Edinburgh, Joseph Black Building, Edinburgh EH9 3JJ, UK.
Nina
N
Svensen
Juan José
JJ
Díaz-Mochón
Mark
M
Bradley
D001667:Binding, Competitive; D002460:Cell Line; D015342:DNA Probes; D004277:DNA, Single-Stranded; D005434:Flow Cytometry; D006801:Humans; D016023:Integrins / Q000378:metabolism; D008024:Ligands; D046228:Microarray Analysis; D021141:Nucleic Acid Amplification Techniques; D019151:Peptide Library; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D054399:Receptors, CCR6 / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018675:Toxicity Tests
C106311:CCR6 protein, human; D015342:DNA Probes; D004277:DNA, Single-Stranded; D016023:Integrins; D008024:Ligands; D019151:Peptide Library; D020135:Peptide Nucleic Acids; D054399:Receptors, CCR6; D011956:Receptors, Cell Surface