| Title: | Responses of the Pheromone-Binding Protein of the Silk Moth Bombyx mori on a Graphene Biosensor Match Binding Constants in Solution |
| Author(s): | Bonazza C; Zhu J; Hasler R; Mastrogiacomo R; Pelosi P; Knoll W; |
| Address: | "CEST Competence Centre for Electrochemical Surface Technology, 2700 Wiener Neustadt, Austria. Austrian Institute of Technology GmbH, Biosensor Technologies, 3430 Tulln, Austria. Department of Agriculture, Food, and Environment, University of Pisa, 56124 Pisa, Italy" |
| ISSN/ISBN: | 1424-8220 (Electronic) 1424-8220 (Linking) |
| Abstract: | "An electronic biosensor for odors was assembled by immobilizing the silk moth Bombyx mori pheromone binding protein (BmorPBP1) on a reduced graphene oxide surface of a field-effect transistor. At physiological pH, the sensor detects the B. mori pheromones, bombykol and bombykal, with good affinity and specificity. Among the other odorants tested, only eugenol elicited a strong signal, while terpenoids and other odorants (linalool, geraniol, isoamyl acetate, and 2-isobutyl-3-methoxypyrazine) produced only very weak responses. Parallel binding assays were performed with the same protein and the same ligands, using the common fluorescence approach adopted for similar proteins. The results are in good agreement with the sensor's responses: bombykol and bombykal, together with eugenol, proved to be strong ligands, while the other compounds showed only poor affinity. When tested at pH 4, the protein failed to bind bombykol both in solution and when immobilized on the sensor. This result further indicates that the BmorPBP1 retains its full activity when immobilized on a surface, including the conformational change observed in acidic conditions. The good agreement between fluorescence assays and sensor responses suggests that ligand-binding assays in solution can be used to screen mutants of a binding protein when selecting the best form to be immobilized on a biosensor" |
| Keywords: | Alkadienes/analysis Biosensing Techniques/*instrumentation/methods Eugenol/analysis Fatty Alcohols/analysis Fluorescence Graphite/chemistry Hydrogen-Ion Concentration Immobilized Proteins/chemistry/*metabolism Insect Proteins/*chemistry/*metabolism Interc; |
| Notes: | "MedlineBonazza, Caroline Zhu, Jiao Hasler, Roger Mastrogiacomo, Rosa Pelosi, Paolo Knoll, Wolfgang eng Switzerland 2021/01/16 Sensors (Basel). 2021 Jan 12; 21(2):499. doi: 10.3390/s21020499" |
