Synthesis and biological activity of a photoaffinity-biotinylated pheromone-biosynthesis activating neuropeptide (PBAN) analog

To study the mode of action of pheromone-biosynthesis activating neuropepti de (PBAN) at the receptor level and for receptor purification, we synthesiz ed and tested the biologic properties of a photoaffinity biotinylated PBAN analog N-[N-(4-azido-tetrafluorobenzoyl)-biocytinyloxyl-succinimide (Atf-Bc t-NHS-PBAN). The Atf-Bct-NHS-PBAN was separated from unreacted reagent and synthetic Hez-PBAN by high-performance liquid chromatography. Conjugated bi otin was detected by using enzyme-linked assay as well as tricine sodium do decyl sulfate polyacrylamide gel electrophoresis. The biologic activity of purified Atf-Bct-NHS-PBAN was confirmed using both in vivo and in vitro phe romonotropic bioassays. These observations indicate that Atf-Bct-NHS-PBAN i s a full agonist of PBAN action in pheromone glands and may be used to stud y PBAN receptors by employing avidin coupled to various reporter groups. (C ) 1999 Elsevier Science Inc. All rights reserved.