Single Step Purification of Locust Adipokinetic Hormones I and II by Reversed-Phase High-Performance Liquid Chromatography, and Amino-Acid Composition of the Hormone II

GÄde, Gerd; Goldsworthy, G.J.; Kegel, Günter; Keller, Rainer

doi:10.1515/bchm2.1984.365.1.393

Cited by 127 publications

(35 citation statements)

References 17 publications

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“…The latter two species were maintained in our insectary in Dusseldorf at 25°C with a 14 h-dark/10 h light cycle and were fed as described previously [5]. [20], and the preparation of methanolic extracts was as in [22].…”

Section: Materials and Methods Insectsmentioning

confidence: 99%

Isolation and structure of a novel charged member of the red–pigment-concentrating hormone-adipokinetic hormone family of peptides isolated from the corpora cardiaca of the blowfly Phormia terraenovae (Diptera)

GÄde

Wilps

Kellner³

1990

Biochemical Journal

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A hypertrehalosaemic neuropeptide from the corpora cardiaca of the blowfly Phormia terraenovae has been isolated by reversed-phase h.p.l.c., and its primary structure was determined by pulsed-liquid phase sequencing employing Edman chemistry after enzymically deblocking the N-terminal pyroglutamate residue. The C-terminus was also blocked, as indicated by the lack of digestion when the peptide was incubated with carboxypeptidase A. The octapeptide has the sequence pGlu-Leu-Thr-Phe-Ser-Pro-Asp-Trp-NH2 and is clearly defined as a novel member of the RPCH/AKH (redpigment-concentrating hormone/adipokinetic hormone) family of peptides. It is the first charged member of this family to be found. The synthetic peptide causes an increase in the haemolymph carbohydrate concentration in a dose-dependent fashion in blowflies and therefore is named ' Phormia terraenovae hypertrehalosaemic hormone' (Pht-HrTH). In addition, receptors in the fat-body of the American cockroach (Periplaneta americana) recognize the peptide, resulting in carbohydrate elevation in the blood. However, fat-body receptors of the migratory locust (Locusta migratoria) do not recognize this charged molecule, and thus no lipid mobilization is observed in this species. INTRODUCTIONIn insects, the corpora cardiaca are major neurohaemal organs, which not only store and release neurosecretory products synthesized in the brain, but also contain intrinsic glandular cells producing a variety of bioactive factors affecting, for example, metabolic processes. Many of these factors appear to be peptides involved in the regulation of carbohydrate release, mostly that of trehalose, the major blood sugar of insects [1,2]. The presence of such hypertrehalosaemic factors is inferred in a wide range of insect species [1,3] in which glycogenolysis, phosphorylase activation or the increase of carbohydrates in the haemolymph occur in response to injections of extracts of their own corpora cardiaca, or in which extracts of their corpora cardiaca are hypertrehalosaemic in cockroaches [1,4,5]. Hypertrehalosaemic

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Section: Materials and Methods Insectsmentioning

confidence: 99%

Isolation and structure of a novel charged member of the red–pigment-concentrating hormone-adipokinetic hormone family of peptides isolated from the corpora cardiaca of the blowfly Phormia terraenovae (Diptera)

GÄde

Wilps

Kellner³

1990

Biochemical Journal

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“…Methanolic extracts of CC were prepared as described previously (Gäde et al, 1984). The dried material was either taken up in water for bioassaying (see below) or it was dissolved in 15% acetonitrile containing 0.1% trifluoroacetic acid (TFA) for reversed-phase high performance liquid chromatography (RP-HPLC) using columns and equipment as described previously (Gäde, 1985; see also legend to Fig.…”

Section: Isolation Of Neuropeptides and Structural Analysesmentioning

confidence: 99%

Bladder grasshoppers (Caelifera: Pneumoridae) contain three adipokinetic peptides

GÄde¹

2008

Eur. J. Entomol.

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Abstract. The corpora cardiaca (CC) of the pneumorid grasshopper species Bullacris discolor contain at least one substance that causes hyperlipaemia in the migratory locust. Isolation of neuropeptides belonging to the adipokinetic hormone (AKH) family was achieved by single-step reversed-phase high performance liquid chromatography (RP-HPLC) of CC extracts and monitoring tryptophan fluorescence. The material of the bladder grasshopper showed three distinct fluorescence peaks with adipokinetic activity in the migratory locust. The peptide sequences were identified by Edman degradation after the N-terminal pyroglutamate residue had been cleaved off enzymatically, and the exact peptide masses were determined by matrix-assisted laser desorption/ionisation time-offlight mass spectrometry. Moreover, the assigned peptides were synthesised and natural and synthetic peptides were compared in their behaviour in RP-HPLC. B. discolor stores three AKH peptides in its CC: two of those are octapeptides, Schgr-AKH-II (pELNFSTGWamide) and Peram-CAH-II (pELTFTPNWamide), whereas the third peptide is a decapeptide, Phyle-CC (pELTFTPNWGSamide. The concentration of carbohydrates in the haemolymph of B. discolor is about 3 times higher than the lipid concentration. Upon injection with synthetic Schgr-AKH-II no adipokinetic or hypertrehalosaemic effect was measurable. A literature survey appears to indicate that an active role of these AKH peptides in substrate mobilisation is only overtly displayed in those caeliferan species that undertake well-defined flight periods.

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“…Isolation of neuropeptides and structural analyses Methanolic extracts of CC were prepared as described previously (Gäde et al, 1984). The dried material was either taken up in water for bioassaying (see below) or it was dissolved in 15% acetonitrile containing 0.1% trifluoroacetic acid (TFA) for reversed-phase high performance liquid chromatography (RP-HPLC) using columns and equipment as described previously (Gäde, 1985; see also legend to Fig.…”

Section: Insectsmentioning

confidence: 99%

Unusual complement of three AKH octapeptides in two species of grasshoppers (Caelifera)

GÄde¹

2006

Eur. J. Entomol.

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Abstract. The corpora cardiaca (CC) of the two grasshopper species Zonocerus elegans (Pyrgomorphidae) and Lamarckiana sparrmani (Pamphagidae) contain (a) substance(s) that cause(s) hyperlipaemia in the migratory locust and hypertrehalosaemia in the American cockroach. Isolation of neuropeptides belonging to the adipokinetic hormone (AKH) family was achieved by single-step reversed-phase high performance liquid chromatography of CC extracts from both species and monitoring tryptophan fluorescence. The material of both species showed three distinct fluorescence peaks with adipokinetic activity in the migratory locust. The peptides were identifid by at least two of the following methods: (1) sequencing by Edman degradation, (2) sequencing by tandem fast atom bombardment mass spectrometry, (3) mass determination by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry and (4) co-elution of the native and synthetic peptides. Both species were found to have three AKH peptides stored in the CC, but unlike in other grasshoppers, none of those peptides were decapeptides. In Z. elegans the following three octapeptides occur: Schgr-AKH-II (pELNFSTGWamide), Peram-CAH-II (pELTFTPNWamide) and Phymo-AKH-III (pEINFTPWWamide), whereas L. sparrmani contains the octapeptides Grybi-AKH (pEVNFSTGWamide), Pyrap-AKH (pELNFTPNWamide) and also Phymo-AKH-III. Conspecific bioassays show no adipokinetic and only a weak (not significant) hypertrehalosaemic effect (in the pamphagid grasshopper). Some explanations are offered on the possible role of these peptides in the species investigated by interpreting their life style.

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Single Step Purification of Locust Adipokinetic Hormones I and II by Reversed-Phase High-Performance Liquid Chromatography, and Amino-Acid Composition of the Hormone II

Cited by 127 publications

References 17 publications

Isolation and structure of a novel charged member of the red–pigment-concentrating hormone-adipokinetic hormone family of peptides isolated from the corpora cardiaca of the blowfly Phormia terraenovae (Diptera)

Isolation and structure of a novel charged member of the red–pigment-concentrating hormone-adipokinetic hormone family of peptides isolated from the corpora cardiaca of the blowfly Phormia terraenovae (Diptera)

Bladder grasshoppers (Caelifera: Pneumoridae) contain three adipokinetic peptides

Unusual complement of three AKH octapeptides in two species of grasshoppers (Caelifera)

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