« Previous AbstractGlutathione peroxidase inhibitory assay for electrophilic pollutants in diesel exhaust and tobacco smoke    Next AbstractHeadspace-free setup of in vitro bioassays for the evaluation of volatile disinfection by-products »

Protein Eng Des Sel

Title:		Modulating and evaluating receptor promiscuity through directed evolution and modeling
Author(s):		Stainbrook SC; Yu JS; Reddick MP; Bagheri N; Tyo KEJ;
Address:		"Interdisciplinary Biological Sciences Program, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208, USA. Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208, USA"
Journal Title:		Protein Eng Des Sel
Year:		2017
Volume:		30
Issue:		6
Page Number:		455 - 465
DOI:		10.1093/protein/gzx018
ISSN/ISBN:		1741-0134 (Electronic) 1741-0126 (Print) 1741-0126 (Linking)
Abstract:		"The promiscuity of G-protein-coupled receptors (GPCRs) has broad implications in disease, pharmacology and biosensing. Promiscuity is a particularly crucial consideration for protein engineering, where the ability to modulate and model promiscuity is essential for developing desirable proteins. Here, we present methodologies for (i) modifying GPCR promiscuity using directed evolution and (ii) predicting receptor response and identifying important peptide features using quantitative structure-activity relationship models and grouping-exhaustive feature selection. We apply these methodologies to the yeast pheromone receptor Ste2 and its native ligand alpha-factor. Using directed evolution, we created Ste2 mutants with altered specificity toward a library of alpha-factor variants. We then used the Vectors of Hydrophobic, Steric, and Electronic properties and partial least squares regression to characterize receptor-ligand interactions, identify important ligand positions and properties, and predict receptor response to novel ligands. Together, directed evolution and computational analysis enable the control and evaluation of GPCR promiscuity. These approaches should be broadly useful for the study and engineering of GPCRs and other protein-small molecule interactions"
Keywords:		"Binding Sites/genetics Directed Molecular Evolution/*methods Least-Squares Analysis *Models, Molecular Protein Binding/genetics Protein Engineering/*methods *Receptors, Cell Surface/chemistry/genetics/metabolism Receptors, Mating Factor/chemistry/genetics;"
Notes:		"MedlineStainbrook, Sarah C Yu, Jessica S Reddick, Michael P Bagheri, Neda Tyo, Keith E J eng S10 OD011996/OD/NIH HHS/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. England 2017/04/30 Protein Eng Des Sel. 2017 Jun 1; 30(6):455-465. doi: 10.1093/protein/gzx018"