Biochemical, Molecular, and Functional Characterization of Piscf-allatostatin, a Regulator of Juvenile Hormone Biosynthesis in the Mosquito Aedes Aegypti.
From: Department of Biological Sciences, Florida International University, Miami, Florida 33199, USA.
The Journal of biological chemistry
- Publish Date: Nov 2006
- ISSN: 0021-9258
- Volume: 281
- Issue: 45
- Pages: 34048-55
- Medium: Print
- Language: English
- Citation (JAMA): Li Yiping, Hernandez-Martinez Salvador, Fernandez Facundo, et al. Biochemical, Molecular, and Functional Characterization of Piscf-allatostatin, a Regulator of Juvenile Hormone Biosynthesis in the Mosquito Aedes Aegypti.. J. Biol. Chem. Nov 2006;281:34048-55
Abstract
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.
Mesh Headings (Keywords): Aedes, Amino Acid Sequence, Animals, Chromatography, High Pressure Liquid, Cloning, Molecular, Corpora Allata, Enzyme-Linked Immunosorbent Assay, Female, Hormone Antagonists, Insect Hormones, Juvenile Hormones, Molecular Sequence Data, Neuropeptides, Peptide Fragments, Polymerase Chain Reaction, Rabbits, Sequence Homology, Amino Acid, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Check for Full Text / PubMed Unique Identifier (PMID): 16968697
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