Reference - Detail
| 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 |
| WOS Category | BIOCHEMISTRY & MOLECULAR BIOLOGY |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 3 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | CHO-K1(RCB0285) |