RRC ID 43635
Author Doi N, Yamakawa N, Matsumoto H, Yamamoto Y, Nagano T, Matsumura N, Horisawa K, Yanagawa H.
Title DNA display selection of peptide ligands for a full-length human G protein-coupled receptor on CHO-K1 cells.
Journal PLoS One
Abstract The G protein-coupled receptors (GPCRs), which form the largest group of transmembrane proteins involved in signal transduction, are major targets of currently available drugs. Thus, the search for cognate and surrogate peptide ligands for GPCRs is of both basic and therapeutic interest. Here we describe the application of an in vitro DNA display technology to screening libraries of peptide ligands for full-length GPCRs expressed on whole cells. We used human angiotensin II (Ang II) type-1 receptor (hAT1R) as a model GPCR. Under improved selection conditions using hAT1R-expressing Chinese hamster ovary (CHO)-K1 cells as bait, we confirmed that Ang II gene could be enriched more than 10,000-fold after four rounds of selection. Further, we successfully selected diverse Ang II-like peptides from randomized peptide libraries. The results provide more precise information on the sequence-function relationships of hAT1R ligands than can be obtained by conventional alanine-scanning mutagenesis. Completely in vitro DNA display can overcome the limitations of current display technologies and is expected to prove widely useful for screening diverse libraries of mutant peptide and protein ligands for receptors that can be expressed functionally on the surface of CHO-K1 cells.
Volume 7(1)
Pages e30084
Published 2012-1-1
DOI 10.1371/journal.pone.0030084
PII PONE-D-11-15339
PMID 22253889
PMC PMC3254644
MeSH Amino Acid Sequence Angiotensin II / chemistry Angiotensin II / genetics Animals Base Sequence Binding, Competitive CHO Cells Calcium / metabolism Cricetinae Cricetulus DNA / metabolism* Humans Ligands Molecular Sequence Data Peptide Library* Peptides / chemistry Peptides / metabolism* Receptor, Angiotensin, Type 1 / metabolism* Recombinant Proteins / metabolism
IF 2.74
Times Cited 4
Human and Animal Cells CHO-K1(RCB0285)