Synthesis and biological activity of a photoaffinity-biotinylated pheromone-biosynthesis activating neuropeptide (PBAN) analog.
To study the mode of action of pheromone-biosynthesis activating neuropeptide (PBAN) at the receptor level and for receptor purification, we synthesized and tested the biologic properties of a photoaffinity biotinylated PBAN analog N-[N-(4-azido-tetrafluorobenzoyl)-biocytinyloxyl-succinimide (Atf-Bct-NHS-PBAN). The Atf-Bct-NHS-PBAN was separated from unreacted reagent and synthetic Hez-PBAN by high-performance liquid chromatography. Conjugated biotin was detected by using enzyme-linked assay as well as tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis. The biologic activity of purified Atf-Bct-NHS-PBAN was confirmed using both in vivo and in vitro pheromonotropic bioassays. These observations indicate that Atf-Bct-NHS-PBAN is a full agonist of PBAN action in pheromone glands and may be used to study PBAN receptors by employing avidin coupled to various reporter groups.
Rafaeli
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A
Department of Stored Products, Volcani Center, Bet Dagan, Israel. vtada@volcani.agri.gov.il
Gileadi
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C
D000345:Affinity Labels / Q000138:chemical synthesis*; D000818:Animals; D001360:Avidin; D001681:Biological Assay; D001710:Biotin; D005260:Female; D066298:In Vitro Techniques; D008297:Male; D009036:Moths; D009479:Neuropeptides / Q000138:chemical synthesis* / Q000494:pharmacology*; D010675:Pheromones / Q000096:biosynthesis*; D018013:Receptors, Neuropeptide / Q000187:drug effects
D000345:Affinity Labels; C400808:N-N-(4-azido-tetrafluorobenzoyl)-biocytinyloxyl-succinimide; D009479:Neuropeptides; D010675:Pheromones; D018013:Receptors, Neuropeptide; D001360:Avidin; D001710:Biotin