Abstract
Novel members of the adipokinetic hormone family of peptides have been identified from the corpora cardiaca (CC) of two species of beetles representing two families, the Silphidae and the Coccinellidae. A crude CC extract (0.3 gland equivalents) of the burying beetle, Nicrophorus vespilloides, was active in mobilizing trehalose in a heterologous assay using the cockroach Periplaneta americana, whereas the CC extract (0.5 gland equivalents) of the ladybird beetle, Harmonia axyridis, exhibited no hypertrehalosemic activity. Primary sequences of one adipokinetic hormone from each species were elucidated by liquid chromatography coupled to electrospray mass spectrometry (LC–MS). The multiple MSN electrospray mass data revealed an octapeptide with an unusual tyrosine residue at position 4 for each species: pGlu-Leu-Thr-Tyr-Ser-Thr-Gly-Trp amide for N. vespilloides (code-named Nicve-AKH) and pGlu-Ile-Asn-Tyr-Ser-Thr-Gly-Trp amide for H. axyridis (code-named Harax-AKH). Assignment of the correct sequences was confirmed by synthesis of the peptides and co-elution in reversed-phase high-performance liquid chromatography with fluorescence detection or by LC–MS. Moreover, synthetic peptides were shown to be active in the heterologous cockroach assay system, but Harax-AKH only at a dose of 30 pmol, which explains the negative result with the crude CC extract. It appears that the tyrosine residue at position 4 can be used as a diagnostic feature for certain beetle adipokinetic peptides, because this feature has not been found in another order other than Coleoptera.
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Acknowledgments
This work was financially supported by funds from the National Research Foundation [Pretoria, South Africa; Grant No. 85768 (IFR13020116790) to GG and IFR2011033100049 to HGM], by staff awards from the Research Council of the University of Cape Town (to GG and HGM) and funds from the Czech Science Foundation (No. 13-18509S to PS). We thank Dr Sandra Steiger (Institute of Experimental Ecology, University of Ulm, Germany), Mr Mike Alsopp (Agricultural Research Council in Stellenbosch, South Africa) and Ms Ingrid A. Minnaar (Center of Excellence for Invasion Biology, University of Stellenbosch, S. Africa) for supplying us with insects. We also acknowledge the assistance of Ms Alukhanyo Xonti in some of the bioassays.
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The authors declare that they have no conflict of interest.
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All applicable international, national and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.
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Gäde, G., Šimek, P. & Marco, H.G. Two novel tyrosine-containing peptides (Tyr4) of the adipokinetic hormone family in beetles of the families Coccinellidae and Silphidae. Amino Acids 47, 2323–2333 (2015). https://doi.org/10.1007/s00726-015-2011-4
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DOI: https://doi.org/10.1007/s00726-015-2011-4