Abstract
Nineteen species of various families of the order Diptera and one species from the order Mecoptera are investigated with mass spectrometry for the presence and primary structure of putative adipokinetic hormones (AKHs). Additionally, the peptide structure of putative AKHs in other Diptera are deduced from data mining of publicly available genomic or transcriptomic data. The study aims to demonstrate the structural biodiversity of AKHs in this insect order and also possible evolutionary trends. Sequence analysis of AKHs is achieved by liquid chromatography coupled to mass spectrometry. The corpora cardiaca of almost all dipteran species contain AKH octapeptides, a decapeptide is an exception found only in one species. In general, the dipteran AKHs are order-specific- they are not found in any other insect order with two exceptions only. Four novel AKHs are revealed by mass spectrometry: two in the basal infraorder of Tipulomorpha and two in the brachyceran family Syrphidae. Data mining revealed another four novel AKHs: one in various species of the infraorder Culicumorpha, one in the brachyceran superfamily Asiloidea, one in the family Diopsidae and in a Drosophilidae species, and the last of the novel AKHs is found in yet another Drosophila. In general, there is quite a biodiversity in the lower Diptera, whereas the majority of the cyclorraphan Brachycera produce the octapeptide Phote-HrTH. A hypothetical molecular peptide evolution of dipteran AKHs is suggested to start with an ancestral AKH, such as Glomo-AKH, from which all other AKHs in Diptera to date can evolve via point mutation of one of the base triplets, with one exception.
Highlights
There are surely a few facts about the importance of Diptera for ecology, medicine and agriculture that are not so well-known to the general public and non-dipteran specialists alike
The remainder of putative adipokinetic hormones (AKHs) sequences were obtained via homology searches using BLAST from the National Center for Biotechnology Information site; AKH peptide precursors from G. morsitans were used as BLAST query
We conducted a conspecific biological assay with only one dipteran species to show proof of principle that the CC extracts of dipteran species do have hypertrehalosemic biological activity
Summary
There are surely a few facts about the importance of Diptera for ecology, medicine and agriculture that are not so well-known to the general public and non-dipteran specialists alike. Regardless of the specific action of these peptides in different insect species, they are generically known as members of the adipokinetic hormone family, in short AKHs. The dipteran AKH was elucidated a year after the tabanid peptides, isolated and sequenced from CC of the blowfly Phormia (=Protophormia) terraenovae (Family: Calliphoridae) as an octapeptide [pELTFSPDW amide; [21]] that regulates carbohydrate metabolism and is released during induced stress [22] and was, code-named Phote-HrTH. The dipteran AKH was elucidated a year after the tabanid peptides, isolated and sequenced from CC of the blowfly Phormia (=Protophormia) terraenovae (Family: Calliphoridae) as an octapeptide [pELTFSPDW amide; [21]] that regulates carbohydrate metabolism and is released during induced stress [22] and was, code-named Phote-HrTH This was the first member of the AKH family that had a charged amino acid; all other AKHs up to were neutral molecules. An additional aim of the current study is to provide a comprehensive list of putative AKH sequences from Diptera by mining publicly-accessible data bases
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