Detection of a Peptide Biomarker by Engineered Yeast Receptors

• DOI: 10.1021/acssynbio.7b00410
• Publication Date: 2018-01-25T02:13:11Z
• Authors: Adebola Adeniran; Sarah Stainbrook; John W. Bostick; Keith E. J. Tyo
• Keywords: Target peptide

Directed evolution of membrane receptors is challenging as the evolved receptor must not only accommodate a non-native ligand, but also maintain ability to transduce detection new ligand any associated intracellular components. The G-protein coupled (GPCR) superfamily largest group receptors. As members GPCR family detect wide range ligands, GPCRs are an incredibly useful starting point for directed user-defined analytical tools and diagnostics. aim this study was determine if yeast Ste2p GPCR, which natively detects α-factor peptide, could yield that Cystatin C, human peptide biomarker. We demonstrate generalizable approach evolving sequences. Because target differs significantly from α-factor, single evolutionary step infeasible. turned substrate walking series chimeric intermediates with increasing similarity validate our previous model tool designing optimal steps. Finally, we clinical utility yeast-based biosensors by showing specific activation C-terminally amidated C in commercially sourced urine. To knowledge, first GPCR.