Autor(es): Li, Yiping; Hernandez-Martinez, Salvador; Fernandez, Facundo; Mayoral, Jaime G.; Topalis, Pantelis; Priestap, Horacio; Perez, Mario; Navare, Arti; Noriega, Fernando G.

Resumo: 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.

Palavras-Chave: Corpora allata; Manduca - Sexta; Iii Biosynthesis; Identification; Allatotropin; Cockroach; Neuropeptides; Genomics; Peptide; Pathway

Imprenta: Journal of Biological Chemistry, v. 281, n. 45, p. 34048-34055, 2006

Identificador do objeto digital: 10.1074/jbc.M606341200

Descritores: Aedes aegypti - Biochemistry ; Aedes aegypti - Biosynthesis ; Aedes aegypti - DNA

Data de publicação: 2006

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