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Invertebrate Neuroscience

, Volume 4, Issue 1, pp 0033–0040 | Cite as

Characterization of a putative pheromone biosynthesis-activating neuropeptide (PBAN) receptor from the pheromone gland of Heliothis peltigera

  • Miriam Altstein
  • Tal Gabay
  • Orna Ben-Aziz
  • Shai Daniel
  • Irina Zeltser
  • Chaim Gilon
ORIGINAL ARTICLE
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Abstract

The binding of [3H]tyrosyl-PBAN28-33NH2 to pheromone gland membranes of the moth Heliothis peltigera was investigated. The study describes the development of a pheromone biosynthesis-activating neuropeptide (PBAN) radioreceptor assay and demonstrates the presence of a putative PBAN binding site on the pheromone gland. It also describes synthesis of a radioligand and optimization of binding conditions with respect to membrane preparation, number of gland equivalents, kinetics of ligand binding and composition of the binding solution. Binding was found to be optimal when membranes were freshly prepared from frozen glands, incubated at a concentration of one gland equivalent per reaction tube in the presence of 10 mM HCO3 ions. Equilibrium of ligand binding was obtained after 20 min. Presence of other components such as NaCl, KCl or SH reagents did not have any effect on binding. Binding was found to be saturable, with a Kd of 5.73 ± 1.05 × 10−6 M and a Bmax of 1.85 ± 0.22 nmol/mg protein. Binding was effectively displaced by unlabeled PBAN1-33NH2 and PBAN28-33ΝΗ2 with a Ki of 4.3 ± 1.1 × 10−6 M and 4.9 ± 2.6 × 10−6 M, respectively.

Key words PBAN, Pheromone biosynthesis-activating neuropeptide receptor Pheromone gland Heliothis peltigera Insect neuropeptide Moths 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Miriam Altstein
    • 1
  • Tal Gabay
    • 1
  • Orna Ben-Aziz
    • 1
  • Shai Daniel
    • 1
  • Irina Zeltser
    • 2
  • Chaim Gilon
    • 2
  1. 1.Department of Entomology, ARO, The Volcani Center, Bet Dagan 50250, Israel e-mail: vinnie2@netvision.net.il Tel.: +972-3-9683710, Fax: +972-3-9683679IL
  2. 2.Department of Organic Chemistry, The Hebrew Univ. of Jerusalem, Jerusalem 91904, IsraelIL

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