Elsevier

Peptides

Volume 20, Issue 1, January 1999, Pages 31-38
Peptides

Original Articles
Isolation and identification of three RFamide-immunoreactive peptides from the mosquito Aedes aegypti

https://doi.org/10.1016/S0196-9781(98)00153-3Get rights and content

Abstract

Three RFamide-immunoreactive peptides were isolated from headless Aedes aegypti mosquitoes. Their structures were identified by Edman degradation or a combination of amino acid composition analysis and mass spectrometry to be: Aedes head peptide 1 [23], [Pro4]-Aedes head peptide (i.e., pGlu-Arg-Pro-Pro-Ser-Leu-Lys-Thr- Arg-Phe-amide), and Leu-Lys-Thr-Arg-Phe-amide which have been named Aedes head peptides 3 and 4, respectively.

Section snippets

Insects

Headless A. aegypti (UGAL-strain) mosquitoes used for peptide isolation were a generous gift from Dr. Mark Brown (Department of Entomology, The University of Georgia, Athens, GA).

ELISA

The competitive RFamide ELISA has been described before in detail [35], it uses a rabbit antiserum to RFamide [15], kindly provided by Dr. Grimmelikhuijzen (now at Copenhagen, Denmark), following the protocol as originally described by Kingan [20] with slight modifications as described elsewhere 33, 35. During

Isolation and identification of Aedes head peptide 3

RFamide immunoreactivity eluted in various fractions from the C18 column. A particular strong peak eluted between 64 and 65 min, and when this material was injected on the C1 column, most of the immunoreactivity eluted between 68 and 71 min, however a smaller amount eluted between 63 and 65 min, and was later shown to contain Aedes head peptide 1 (see below). After injection on the microsorb phenyl column a large UV-absorbing and RFamide-immunoreactive peak was found to elute at 46.0 min (Fig. 1

Discussion

Aedes head peptide 1 was the first peptide identified of what now appears to be an arthropod peptide family (Table 3 ). Its members have been identified from the horse shoe crab, Limulus polyphemus [38], the American cockroach, Periplaneta americana [35], the mosquito A. aegypti [23], and the Colorado potato beetle, Leptinotarsa decemlineata [32]. It has been reported, that the Aedes head peptide 1 inhibits host-seeking behavior during oogenesis [8] and it is of interest to note that the same

Acknowledgements

I thank Dr. Mark R. Brown for his generous gift of headless mosquitoes, Dr. C. J. P. Grimmelikhuijzen for an aliquot of his RFamide antiserum, Dr. Thomas McClure for mass spectrometry, Mr. Wallace Clark for amino acid composition and sequence analysis, Dr. Ron Jasensky for peptide synthesis, and Dr. Henry H. Hagedorn for use of laboratory space. This work was supported by grants from the National Institutes of Health (AI-33429) to the author and from the John D. and Catherine T. McArthur

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