Biochemical, molecular, and functional characterization of PISCF-allatostatin, a regulator of juvenile hormone biosynthesis in the mosquito Aedes aegypti

J Biol Chem. 2006 Nov 10;281(45):34048-55. doi: 10.1074/jbc.M606341200. Epub 2006 Sep 12.


Aedes aegypti PISCF-allatostatin or allatostatin-C (Ae-AS-C) was isolated using a combination of high performance liquid chromatography and enzyme-linked immunosorbent assay (ELISA). The matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrum of positive ELISA fractions revealed a molecular mass of 1919.0 Da, in agreement with the sequence qIRYRQCYFNPISCF, with bridged cysteines. This sequence was confirmed by matrix-assisted laser desorption/ionization tandem TOF/TOF mass spectrometry analysis. The corresponding Ae-AS-C cDNA was amplified by PCR, and the sequence of the peptide was confirmed. An in vitro radiochemical assay was used to study the inhibitory effect of synthetic Ae-AS-C on juvenile hormone biosynthesis by the isolated corpora allata (CA) of adult female A. aegypti. The inhibitory action of synthetic Ae-AS-C was dose-dependent; with a maximum at 10(-9) m. Ae-AS-C showed no inhibitory activity in the presence of farnesoic acid, an immediate precursor of juvenile hormone, indicating that the Ae-AS-C target is located before the formation of farnesoic acid in the pathway. The sensitivity of the CA to inhibition by Ae-AS-C in the in vitro assay varied during the adult life; the CA was most sensitive during periods of low synthetic activity. In addition, the levels of Ae-AS-C in the brain were studied using ELISA and reached a maximum at 3 days after eclosion. These studies suggest that Ae-AS-C is an important regulator of CA activity in A. aegypti.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Aedes / drug effects
  • Aedes / growth & development
  • Aedes / metabolism*
  • Amino Acid Sequence
  • Animals
  • Chromatography, High Pressure Liquid
  • Cloning, Molecular
  • Corpora Allata / drug effects*
  • Corpora Allata / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Hormone Antagonists / chemical synthesis
  • Hormone Antagonists / chemistry
  • Hormone Antagonists / pharmacology*
  • Insect Hormones / biosynthesis*
  • Insect Hormones / genetics
  • Juvenile Hormones / biosynthesis*
  • Juvenile Hormones / genetics
  • Molecular Sequence Data
  • Neuropeptides / chemical synthesis
  • Neuropeptides / chemistry
  • Neuropeptides / pharmacology*
  • Peptide Fragments / immunology
  • Peptide Fragments / metabolism
  • Polymerase Chain Reaction
  • Rabbits
  • Sequence Homology, Amino Acid
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization


  • Hormone Antagonists
  • Insect Hormones
  • Juvenile Hormones
  • Neuropeptides
  • Peptide Fragments
  • allatostatin