Lepidoptera is amongst one of the four most speciose insect orders and ecologically very successful because of their ability to fly. Insect flight is always aerobic and exacts a high metabolic demand on the animal. A family of structurally related neuropeptides, generically referred to as adipokinetic hormones (AKHs), play a key role in triggering the release of readily utilizable fuel metabolites into the hemolymph from the storage forms in the fat body. We used mass spectrometry to elucidate AKH sequences from 34 species of Lepidoptera and searched the literature and publicly available databases to compile (in a phylogenetic context) a comprehensive list of all Lepidoptera sequences published/predicted from a total of 76 species. We then used the resulting set of 15 biochemically characterized AKHs in a physiological assay that measures lipid or carbohydrate mobilization in three different lepidopteran species to learn about the functional cross-activity (receptor-ligand interactions) amongst the different butterfly/moth families. Our results include novel peptide structures, demonstrate structural diversity, phylogenetic trends in peptide distribution and order-specificity of Lepidoptera AKHs. There is almost an equal occurrence of octa-, nona-, and decapeptides, with an unparalleled emphasis on nonapeptides than in any insect order. Primitive species make Peram-CAH-II, an octapeptide found also in other orders; the lepidopteran signature peptide is Manse-AKH. Not all of the 15 tested AKHs are active in Pieris brassicae; this provides insight into structure-activity specificity and could be useful for further investigations into possible biorational insecticide development.
- Publikační typ
- časopisecké články MeSH
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.
- MeSH
- chromatografie kapalinová MeSH
- Diptera chemie klasifikace genetika metabolismus MeSH
- hmotnostní spektrometrie MeSH
- hmyzí hormony analýza chemie genetika metabolismus MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty analýza chemie metabolismus MeSH
- molekulární evoluce * MeSH
- oligopeptidy analýza chemie genetika metabolismus MeSH
- peptidy analýza chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- vztahy mezi strukturou a aktivitou MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Seventeen species of the coleopteran series Cucujiformia are investigated for the presence and sequence of putative adipokinetic hormones (AKHs). Cucujiformia includes species from the major superfamilies, that is, Chrysomeloidea, Curculionoidea, Cucujoidea, and Tenebrionoidea. The clade Phytophaga in which the Chrysomeloidea and Curculionoidea reside, harbor very detrimental species for agriculture and forestry. Thus, this study aims not only to demonstrate the structural biodiversity of AKHs in these beetle species and possible evolutionary trends but also to determine whether the AKHs from harmful pest species can be used as lead substances for a future putative insecticide that is harmless to beneficial insects. Sequence analysis of AKHs is achieved by liquid chromatography coupled to mass spectrometry. Most of the investigated species contain AKH octapeptides in their corpora cardiaca, although previously published work also found a few decapeptides, which we comment on. The signature and sole AKH in cerambycidae Chrysomeloidea and Curculionoidea is Peram-CAH-I (pEVNFSPNW amide), which is also found in the majority of chrysomelidae Chrysomeloidea and in the one investigated species of Cucujoidea albeit in a few cases associated with a second AKH which can be either Peram-CAH-II (pELTFTPNW amide), Emppe-AKH (pEVNFTPNW amide), or Micvi-CC (pEINFTPNW amide). The most often encountered AKH in Tenebrionoidea, family Meloidae as well as family Tenebrionidae, is Tenmo-HrTH (pELNFSPNW amide) followed by Pyrap-AKH (pELNFTPNW amide) and a Tenmo-HrTH extended decapeptide (in Meloidae). Finally, we examine AKH sequences from 43 species of cucujiform beetles, including the superfamily Coccinelloidea for a possible lead compound for producing a cucujiform-specific pesticide.
- MeSH
- brouci chemie MeSH
- chromatografie kapalinová MeSH
- corpora allata chemie MeSH
- hmotnostní spektrometrie MeSH
- hmyzí hormony analýza chemie MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty analýza chemie MeSH
- oligopeptidy analýza chemie MeSH
- peptidy analýza chemie MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The rationale of "green pesticides" in food security is to use information about endogenous hormones of pest insects to make peptide mimetics that will act against the pest insects to alter their behaviour or physiology, while taking care not to harm beneficial insects or other organisms in the food chain. Such "green" insecticides are designed thus, on the basis of neuropeptide ligand-receptor interaction and it is of paramount interest to have finally a mimetic at hand that is harmful only to pest insects. For this concept to work, one has to identify the ligands in pest and beneficial insects. In this study we investigate adipokinetic hormones (AKHs) from a hemipteran source. The most harmful hemipterans on an economic scale are aphids (Hemiptera: Sternorrhyncha: Aphidoidea) of which the AKH is known. Here we identify the AKH complement of a member of a related suborder, the raintree bug or froghopper Ptyelus flavescens (Hemiptera: Cicadomorpha: Cercopoidea). Identification and sequence elucidation of the adipokinetic peptides of this species was achieved by a heterospecific and conspecific trehalose-mobilizing bioassay, and by liquid chromatography coupled to positive electrospray mass spectrometry (LC-ESI-MS) including tandem MS2spectra obtained by collision-induced dissociation. High resolution MS was employed to distinguish between Gln and Lys residues in the peptides. Three AKHs are discovered in the raintree bug: an octapeptide (Peram-CAH-I: pEVNFSPNW amide) previously known from cockroaches, and two novel decapeptides (Ptyfl-AKH-I: pEINFSTGWGQ amide and Ptyfl-AKH-II: pEINFSTAWGQ amide). The novel peptides were synthesized and the sequence assignments were unequivocally confirmed by co-elution of synthetic peptides and the natural equivalent and by identical MS data of the two forms. A conspecific bioassay in the froghopper describe the endogenous peptide Ptyfl-AKH-I as hypertrehalosemic. In heterologous bioassays the two novel AKHs induce an increase of circulating carbohydrates in cockroaches: Ptyfl-AKH-I is much more active than Ptyfl-AKH-II. Moreover, if the Ile2in Ptyfl-AKH-II is replaced with a Leu2residue, biological activity is further diminished. The current data show that the raintree AKH decapeptides differ by four amino acids from the aphid AKH (Acypi-AKH: pEVNFTPTWGQ amide). Therefore, it may be permissible to use the aphid AKH ligand-receptor pair to develop a "green" insecticide to target aphid metabolism.
Eight beetle species of the superfamily Scarabaeoidea were investigated with respect to peptides belonging to the adipokinetic hormone (AKH) family in their neurohemal organs, the corpora cardiaca (CC). The following beetle families are represented: Scarabaeidae, Lucanidae, and Geotrupidae. AKH peptides were identified through a heterospecific trehalose-mobilizing bioassay and by sequence analyses, using liquid chromatography coupled to positive electrospray mass spectrometry (LC-ESI-MS) and analysis of the tandem MS2 spectra obtained by collision-induced dissociation. All the beetle species have octapeptide AKHs; some have two AKHs, while others have only one. Novel AKH members were found in Euoniticellus intermedius and Circellium bacchus (family Scarabaeidae), as well as in Dorcus parallelipipedus (family Lucanidae). Two species of the family Geotrupidae and two species of the Scarabaeidae subfamily Cetoniinae contain one known AKH peptide, Melme-CC, while E. intermedius produces a novel peptide code named Euoin-AKH: pEINFTTGWamide. Two AKH peptides were each identified in CC of C. bacchus and D. parallelipipedus: the novel Cirba-AKH: pEFNFSAGWamide and the known peptide, Scade-CC-I in the former, and the novel Dorpa-AKH: pEVNYSPVW amide and the known peptide, Melme-CC in the latter. Kheper bonelli (subfamily Scarabaeinae) also has two AKHs, the known Scade-CC-I and Scade-CC-II. All the novel peptides were synthesized and the amino acid sequence assignments were unequivocally confirmed by co-elution of the synthetic peptides with their natural equivalent, and identical MS parameters of the two forms. The novel synthetic peptides are all active in inducing hypertrehalosemia in cockroaches.
- MeSH
- brouci chemie genetika MeSH
- chromatografie kapalinová MeSH
- hmotnostní spektrometrie MeSH
- hmyzí hormony chemie genetika izolace a purifikace MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty chemie izolace a purifikace MeSH
- oligopeptidy chemie genetika izolace a purifikace MeSH
- peptidy chemie genetika izolace a purifikace MeSH
- sekvence aminokyselin genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A species of the poorly studied order Embioptera, the webspinner Oligotoma saundersii, is investigated for its complement of neuropeptides of the adipokinetic hormone (AKH) family. A methanolic extract of its corpora cardiaca (CC) is able to effect carbohydrate mobilization in the cockroach, Periplaneta americana, and liquid chromatography coupled to electrospray ionization mass spectrometry clearly identified one decapeptide as a member of the AKH family in the CC of O. saundersii. The primary structure of this peptide, code-named Olisa-AKH, is elucidated as pEVNFSPNWGG amide. It is a novel member of the AKH family and in its synthetic form it has strong hypertrehalosemic activity in the American cockroach. This effect may be explained by its near-identical structure compared with one of the endogenous cockroach AKH peptides. An analog with the reversed order of the proline and asparagine residues, viz. N(6)P(7)-Olisa-AKH, had negligible activity thus, shedding light on the requirements of the cockroach AKH receptor. From reversed-phase high-performance liquid chromatography experiments, we can conclude that the CC from an individual webspinner contains less than one pmol of Olisa-AKH. Comparison of the AKH sequences from the major orders of the Polyneoptera does not point to a close phylogenetic relationship between webspinners and stick insects.
- MeSH
- hmyzí proteiny * chemie genetika izolace a purifikace MeSH
- neuropeptidy * chemie genetika izolace a purifikace MeSH
- Periplaneta chemie genetika MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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 MS(N) 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.
- MeSH
- adipokiny chemie genetika izolace a purifikace MeSH
- brouci chemie genetika MeSH
- hmyzí hormony chemie genetika izolace a purifikace MeSH
- hmyzí proteiny chemie genetika izolace a purifikace MeSH
- molekulární sekvence - údaje MeSH
- peptidy chemie genetika izolace a purifikace MeSH
- sekvence aminokyselin * MeSH
- tyrosin chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The basal caeliferan family Tetrigidae is investigated to identify neuropeptides belonging to the adipokinetic hormone (AKH) family. The pygmy grasshopper Tetrix subulata contains in its corpus cardiacum two octapeptides as revealed by liquid chromatography coupled to electrospray ionization mass spectrometry. The less abundant peptide is the well-known Schgr-AKH-II (pELNFSTGW amide) which is suggested to be the ancestral AKH of Caelifera and Ensifera. The second peptide, Tetsu-AKH (pEFNFTPGW amide), is novel and quite unusual with its third aromatic residue at position 2. It is thought to be autapomorphic for Caelifera. Tetsu-AKH has hyperlipemic activity in T. subulata and in Schistocerca gregaria.
- MeSH
- adipokiny chemie izolace a purifikace farmakologie MeSH
- corpora allata metabolismus MeSH
- hmyzí proteiny chemie izolace a purifikace farmakologie MeSH
- kobylky chemie MeSH
- metabolismus lipidů účinky léků MeSH
- molekulová hmotnost MeSH
- sekvence aminokyselin MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Two extant apterygotan insect orders, Archaeognatha and Zygentoma, are investigated with respect to the identity of neuropeptides belonging to the adipokinetic hormone (AKH) peptide family; this is the first report on AKH peptide structures in the so-called primitive insects and the first of any peptide in the Archaeognatha. In the lepismatid, Thermobia domestica, and the machilid, Petrobius maritimus, a single AKH peptide is identified and sequenced from each species; neither sequence is novel and has previously been shown in corpora cardiaca (CC) of cockroaches (Peram-CAH-I) and dragonflies (Anaim-AKH), respectively. These octapeptides differ from each other only in one position (Asn(7) in the lepismatid and Ser(7) in the machilid). The biological relevance of these peptides was investigated and we speculate that they are likely involved in the mobilisation of lipids in the apterygotes. Immunocytochemistry with an antibody directed against an AKH revealed a well-developed pair of CC in T. domestica and another lepismatid, the fishmoth Ctenolepisma longicaudata; a cluster of immunopositive cells are located retrocerebrally in tissue sections of P. maritimus which may be the CC.
- MeSH
- chromatografie kapalinová MeSH
- hmotnostní spektrometrie s elektrosprejovou ionizací MeSH
- hmyz metabolismus MeSH
- hmyzí hormony metabolismus sekrece MeSH
- imunohistochemie MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty metabolismus MeSH
- metabolismus lipidů MeSH
- oligopeptidy metabolismus sekrece MeSH
- trehalosa metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Peptides of the adipokinetic hormone (AKH)/red pigment-concentrating hormone (RPCH) family were isolated and sequenced from the retrocerebral corpora cardiaca of four kissing bugs which are all vectors of the protozoan Trypanosoma cruzi responsible for Chagas' disease. The sequence of three novel AKHs were deduced from the multiple MS(N) electrospray mass data: the octapeptide pGlu-Leu-Thr-Phe-Ser-Thr-Asp-Trp amide (denoted Rhopr-AKH) in Rhodnius prolixus and Panstrongylus megistus, the nonapeptide pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp-Gly amide (denoted Triin-AKH) in Triatoma infestans and the decapeptide pGlu-Leu-Thr-Phe-Ser-Asp-Gly-Trp-Gly-Asn amide (denoted Dipma-AKH) in Dipetalogaster maxima. The sequences were confirmed by identical behavior of natural and synthetic forms in reversed-phase HPLC and by CID-MS mass spectra. Conspecific injections of a dose of 10 pmol of the respective synthetic peptides resulted in a small but significant increase of the lipid concentration in the hemolymph. These experiments suggest that AKHs in kissing bugs act to regulate lipid metabolism, possibly during dispersal flights which is one of the mechanisms whereby the insects reach new outbreak areas. Copyright © 2012 Elsevier Inc. All rights reserved.
- MeSH
- hemolymfa metabolismus MeSH
- hmyzí hormony MeSH
- kobylky metabolismus účinky léků MeSH
- kyselina pyrrolidonkarboxylová * MeSH
- metabolismus lipidů MeSH
- oligopeptidy MeSH
- Panstrongylus * metabolismus MeSH
- Rhodnius * metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční analýza proteinů MeSH
- Triatoma * metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
