Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Isolation and identification of 3-acetylecdysone 2-phosphate, a metabolite of ecdysone, from developing eggs of Schistocerca gregaria.

ABSTRACT: A major ecdysteroid conjugate, which accumulates in the eggs of the desert locust, Schistocerca gregaria, during the later stages of embryogenesis, has been isolated by reversed-phase and anion-exchange high-performance liquid chromatography. Hydrolysis of the conjugate with a crude arylsulphatase preparation from Helix pomatia liberates mainly ecdysone 3-acetate. The compound was identified as 3-acetylecdysone 2-phosphate by phosphate analysis of an acid-hydrolysed sample, fast atom bombardment, electron impact and chemical ionization mass spectrometry and 1H and 13Cn.m.r. spectroscopy. The instability of 3-acetylecdysone 2-phosphate on storage results in the formation of ecdysone 2-phosphate, which was identified by physicochemical techniques. 3-Acetylecdysone 2-phosphate and ecdysone 2-phosphate are less susceptible than ecdysone 22-phosphate to hydrolysis in vitro by an enzyme preparation from S. gregaria embryos. The possible role of 3-acetylecdysone 2-phosphate as an inactive end product of ecdysteroid metabolism is discussed.

SUBMITTER: Isaac RE

PROVIDER: S-EPMC1153201 | biostudies-other | 1984 Jan

REPOSITORIES: biostudies-other

ACCESS DATA

Json Xml

Similar Datasets

Isolation and identification of ecdysteroid phosphates and acetylecdysteroid phosphates from developing eggs of the locust, Schistocerca gregaria.

Project description:Maturing eggs of the desert locust, Schistocerca gregaria, contain a variety of ecdysteroid (insect moulting hormone) conjugates and metabolites, four of which have been previously isolated from polar extracts and identified as ecdysonoic acid, 20-hydroxyecdysonoic acid, 3-acetylecdysone 2-phosphate and ecdysone 2-phosphate. In the present study we have isolated eight additional ecdysteroids from similar late-stage eggs by high-performance liquid chromatography. The 22-phosphate esters of ecdysone, 2-deoxyecdysone, 20-hydroxyecdysone and 2-deoxy-20-hydroxyecdysone, all of which were first identified as ecdysteroid components of newly-laid eggs of S. gregaria, were identified by co-chromatography with authentic compounds and by physicochemical techniques. The remaining compounds were identified as 3-acetyl-20-hydroxyecdysone 2-phosphate, 3-epi-2-deoxyecdysone 3-phosphate, 3-acetylecdysone 22-phosphate and 2-acetylecdysone 22-phosphate by fast atom bombardment mass spectrometry, p.m.r. spectroscopy and analysis of the steroid moieties after enzymic hydrolysis. The latter two compounds, after isolation, are susceptible to nonenzymic acetyl migration and deacetylation to give mixtures of ecdysone 22-phosphate and its 2- and 3-acetate derivatives. The possible role and significance of these ecdysteroid conjugates with respect to the control of hormone titres in insect eggs is discussed.

| S-EPMC1144059 | biostudies-other

Identification of the 22-phosphate esters of ecdysone, 2-deoxyecdysone, 20-hydroxyecdysone and 2-deoxy-20-hydroxyecdysone from newly laid eggs of the desert locust, Schistocerca gregaria.

Project description:The four major ecdysteroid (insect moulting hormone) conjugates present in the newly laid eggs of the desert locust, Schistocera gregaria, have been purified by reversed-phase and anion-exchange high-performance liquid chromatography. The steroid moieties were identified as ecdysone, 2-deoxyecdysone, 20-hydroxyecdysone and 2-deoxy-20-hydroxyecdysone. Phosphate analysis of acid-hydrolysed samples showed a steroid:phosphate ratio of approx. 1:1 for all four compounds. The intact conjugates were identified as ecdysone 22-phosphate, 2-deoxyecdysone 22-phosphate, 20-hydroxyecdysone 22-phosphate and 2-deoxy-20-hydroxyecdysone 22-phosphate by fast atom bombardment mass spectrometry and 1H, 13C and 31P n.m.r. The significance of ecdysteroid phosphates as a source of free hormone during embryogenesis is discussed.

| S-EPMC1152159 | biostudies-other

Identification of ecdysonoic acid and 20-hydroxyecdysonoic acid isolated from developing eggs of Schistocerca gregaria and pupae of Spodoptera littoralis.

Project description:Ecdysonoic acid and 20-hydroxyecdysonoic acid have been purified from developing eggs of the desert locust, Schistocerca gregaria, by high performance liquid chromatography (h.p.l.c.), and their structures were determined by p.m.r. spectroscopy and fast atom bombardment mass spectrometry of the free and methyl ester derivatives. 20-Hydroxyecdysonoic acid was also characterized from Spodoptera littoralis pupae. The occurrence of both 20-hydroxyecdysonoic acid and ecdysonoic acid in Sp. littoralis pupae was also established by h.p.l.c. comparison of the 3H-labelled acids formed from [3H]ecdysone and of their methyl esters with the corresponding substances from Sch. gregaria. The significance of ecdysteroid acids as products of ecdysteroid inactivation is discussed.

| S-EPMC1152117 | biostudies-other

Ecdysone 20-mono-oxygenase in the desert locust, Schistocerca gregaria.

Project description:The enzyme catalysing the hydroxylation of ecdysone to 20-hydroxyecdysone, ecdysone 20-mono-oxygenase (EC 1.14.99.22), was investigated in the Malpighian tubules of fifth-instar locusts, Schistocerca gregaria. Enzyme activity was optimal at 35 degrees C and pH 6.8-8.0. Under these conditions the mono-oxygenase exhibited an apparent Km for ecdysone of 7.1 X 10(-7) M, a maximal specific activity of 1.1 nmol/h per mg of protein and was competitively inhibited by 20-hydroxyecdysone with an apparent Ki of 6.3 X 10(-7) M. Enzyme activity was decreased in the presence of Ca2+, Mg2+, EDTA and non-ionic detergents. The Malpighian tubule ecdysone 20-mono-oxygenase was localized primarily in the subcellular fraction sedimenting at 7500 g and, on the basis of marker enzyme profiles, was assigned mainly to the mitochondria. NADPH was required for activity, although addition of NADH together with NADPH had a synergistic effect. NADP+-dependent isocitrate dehydrogenase (EC 1.1.1.42) and an energy-dependent NAD(P) transhydrogenase (EC 1.6.1.1.) appeared to be the major sources of reducing equivalents, with the contribution from the 'malic enzyme' (EC 1.1.1.40) being less important. The monooxygenase was characterized as a cytochrome P-450-containing mixed-function oxidase from the inhibition patterns with metyrapone, CO and cyanide; CO inhibition was reversible with monochromatic light at 450 nm. However, the ecdysone 20-mono-oxygenase shows much lower sensitivity to CO inhibition and to photodissociation of the CO-inhibited complex than do vertebrate cytochrome P-450-dependent hydroxylation systems. The concentration of cytochrome P-450 in the Malpighian tubule mitochondria was 30 pmol/mg of protein. The properties of the mono-oxygenase are discussed in relation to hydroxylation enzymes from other sources.

| S-EPMC1144370 | biostudies-other

The ecdysone receptor complex is essential for the reproductive success in the female desert locust, Schistocerca gregaria.

Project description:Ecdysteroid hormones influence the development and reproduction of arthropods by binding a heterodimeric complex of nuclear receptors, the ecdysone receptor (EcR) and the retinoid-X-receptor/ultraspiracle (RXR/USP). Here, we report on the in vivo role(s) of the ecdysone receptor complex, SchgrEcR/SchgrRXR, in the female reproductive physiology of a major phytophagous pest insect, i.e. the desert locust, Schistocerca gregaria. Tissue and temporal distribution profiles were analysed during the first gonadotrophic cycle of adult female locusts. RNA interference was used as a reverse genetics tool to investigate the in vivo role of the ecdysone receptor complex in ovarian maturation, oogenesis, fertility and fecundity. We discovered that silencing the ecdysone receptor complex in S. gregaria resulted in impaired ovulation and oviposition, indicative for a crucial role of this complex in chorion formation. We also found evidence for a feedback of SchgrEcR/SchgrRXR on juvenile hormone biosynthesis by the corpora allata. Furthermore, we observed a tissue-dependent effect of the SchgrEcR/SchgrRXR knockdown on the transcript levels of the insulin receptor and neuroparsin 3 and 4. The insulin receptor transcript levels were upregulated in the brain, but not the fat body and gonads. Neuroparsins 3 and 4 transcript levels were down regulated in the brain and fat body, but not in the gonads.

| S-EPMC6327042 | biostudies-literature