Title: | PBAN receptor: employment of anti-receptor antibodies for its characterization and for development of a microplate binding assay |
Author(s): | Ben Yosef T; Bronshtein A; Ben Aziz O; Davidovitch M; Tirosh I; Altstein M; |
Address: | "Department of Entomology, The Volcani Center, Hamabacim St. 6, Bet Dagan 50250, Israel" |
DOI: | 10.1016/j.jinsphys.2009.05.006 |
ISSN/ISBN: | 1879-1611 (Electronic) 0022-1910 (Linking) |
Abstract: | "This study describes generation of an anti-PBAN receptor (PBAN-R) antiserum and its employment for the characterization of the PK/PBAN-R(s). The antiserum recognized, in a specific and dose-dependent manner, the presence of PBAN-R in pheromone gland membrane preparations of three female moths: Heliothis peltigera, Helicoverpa armigera and Spodoptera littoralis. It also reacted specifically with the S. littoralis larval receptor in vivo, most likely by competing with the ligand on the binding site and consequently inhibiting cuticular melanization. Despite its ability to react with the receptor of H. peltigera in dot blot experiments, the antiserum did not react with the receptor in vivo and failed to inhibit sex pheromone biosynthesis. The antiserum was also used to develop two microplate binding assays. The Ab described in this study is the first raised against an insect neuropeptide (Np) receptor to be used in vivo, and its employment for characterization of the PK/PBAN-R(s) may thus provide important information on the mode of action of this Np family. The present study adds important information on the difference between the receptors in the two moth species, hints at the possible existence of receptor subtypes, and provides a platform for the development of a high-throughput assay (HTA) for screening of PK/PBAN agonists and antagonists" |
Keywords: | "Animals Antibodies/*analysis Binding Sites Enzyme-Linked Immunosorbent Assay/*methods Female Insect Proteins/*chemistry/metabolism Moths/*chemistry/metabolism Protein Binding Rabbits Receptors, Neuropeptide/*chemistry/metabolism;" |
Notes: | "MedlineBen Yosef, Tal Bronshtein, Alisa Ben Aziz, Orna Davidovitch, Michael Tirosh, Itay Altstein, Miriam eng Evaluation Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. England 2009/06/02 J Insect Physiol. 2009 Sep; 55(9):825-33. doi: 10.1016/j.jinsphys.2009.05.006. Epub 2009 Jun 10" |