scholarly journals Proctolin degradation by membrane peptidases from nervous tissues of the desert locust (Schistocerca gregaria)

1987 ◽  
Vol 245 (2) ◽  
pp. 365-370 ◽  
Author(s):  
R E Isaac
Keyword(s):  
Schistocerca Gregaria ◽  
Physiological Role ◽  
Hydrolytic Activity ◽  
Mitochondrial Fraction ◽  
Membrane Preparation ◽  
Aminopeptidase Activity ◽  
Synaptic Membrane ◽  
Desert Locust ◽  
Ph Optimum ◽  
Enzyme Preparations

The hydrolysis of the insect neuropeptide proctolin (Arg-Tyr-Leu-Pro-Thr) by enzyme preparations from the nervous tissue of the desert locust (Schistocerca gregaria) was investigated. Neural homogenate degraded proctolin (100 microM) at neutral pH by cleavage of the Arg-Tyr and Tyr-Leu bonds to yield Tyr-Leu-Pro-Thr, Arg-Tyr and free tyrosine. Arg-Tyr was detected as a major metabolite when the aminopeptidase inhibitors amastatin and bestatin were present to prevent Arg-Tyr breakdown. Around 50% of the proctolin-degrading activity was isolated in a 30,000 g membrane fraction and was shown to be almost entirely due to aminopeptidase activity. The aminopeptidase had an apparent Km of 23 microM, a pH optimum of 7.0 and was inhibited by 1 mM-EDTA and amastatin [IC50 = 0.3 microM], but was relatively insensitive to bestatin, actinonin and puromycin. Phenylmethanesulphonyl fluoride (1 mM) and p-chloromercuriphenylsulphonic acid (1 mM) had no effect on this enzyme activity. Although the bulk of the Tyr-Leu hydrolytic activity was located in the 30,000 g supernatant, some weak activity was detected in a washed membrane preparation. This peptidase displayed a high affinity for proctolin (Km = 0.35 microM) and optimal activity at around pH 7.0. Synaptosome- and mitochondria-rich fractions were prepared from crude neural membranes. The aminopeptidase activity was concentrated in the synaptic-membrane preparation, whereas activity giving rise to Arg-Tyr was predominantly localized in the mitochondrial fraction. The subcellular localization of the membrane aminopeptidase is consistent with a possible physiological role for this enzyme in the inactivation of synaptically released proctolin.

scholarly journals Neuropeptide-degrading endopeptidase activity of locust (Schistocerca gregaria) synaptic membranes

1988 ◽  
Vol 255 (3) ◽  
pp. 843-847 ◽  
Author(s):  
R E Isaac
Keyword(s):  
Schistocerca Gregaria ◽  
Physiological Role ◽  
Aminopeptidase Activity ◽  
Synaptic Membranes ◽  
Synaptic Membrane ◽  
Adipokinetic Hormone ◽  
Peptide Bonds ◽  
Endopeptidase 24.11 ◽  
Endopeptidase Activity ◽  
Neural Membranes

Locust adipokinetic hormone (AKH, pGlu-Leu-Asn-Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2) was used as the substrate to measure neuropeptide-degrading endopeptidase activity in neutral membranes from ganglia of the locust Schistocerca gregaria. Initial hydrolysis of AKH at neural pH by peptidases of washed neural membranes generated pGlu-Leu-Asn and Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2 as primary metabolites, demonstrating that degradation was initiated by cleavage of the Asn-Phe bond. Amastatin protected the C-terminal fragment from further metabolism by aminopeptidase activity without inhibiting AKH degradation. The same fragments were generated on incubation of AKH with purified pig kidney endopeptidase 24.11, and enzyme known to cleave peptide bonds that involve the amino group of hydrophobic amino acids. Phosphoramidon (10 microM), a selective inhibitor of mammalian endopeptidase 24.11, partially inhibited the endopeptidase activity of locust neural membranes. This phosphoramidon-sensitive activity was shown to enriched in a synaptic membrane preparation with around 80% of the activity being inhibited by 10 microM-phosphoramidon (IC50 = 0.2 microM). The synaptic endopeptidase was also inhibited by 1 mM-EDTA, 1 mM-1,10-phenanthroline and 1 microM-thiorphan, and the activity was maximal between pH 7.3 and 8.0. Localization of the phosphoramidon-sensitive enzyme in synaptic membranes is consistent with a physiological role for this endopeptidase in the metabolism of insect peptides at the synapse.

scholarly journals The mechanism of C-20 hydroxylation of α-ecdysone in the desert locust, Schistocerca gregaria

1977 ◽  
Vol 168 (3) ◽  
pp. 513-520 ◽  
Author(s):  
P Johnson ◽  
H H Rees
Keyword(s):  
Schistocerca Gregaria ◽  
Fat Body ◽  
Difference Spectrum ◽  
Maximum Activity ◽  
Malpighian Tubules ◽  
Desert Locust ◽  
Ph Optimum ◽  
Developmental Variation ◽  
Moulting Hormone ◽  
Cytochrome P 450

1. The C-20 hydroxylation of alpha-ecdysone to produce beta-ecdysone was investigated in the desert locust, Schistocerca gregaria. 2. alpha-Ecdysone C-20 hydroxylase activity was located primarily in the fat-body and Malpighian tubules. The properties of the hydroxylation system from Malpighian tubules investigated further. 3. The enzyme system was mitochondrial, had a pH optimum of 6.5, an apparent Km of 12.5 micron and required O2 and NADPH. 4. The activity of the hydroxylation system showed developmental variation within the fifth instar, the maximum activity corresponding to the maximum tire of endogenous moulting hormone. The significance of these results is assessed in relation to the control of the endogenous titre of beta-ecdysone. 5. The mechanism of the hydroxylation system was investigated by using known inhibitors of hydroxylation reactions such as CO, metyrapone and cyanide. 6. The CO difference spectrum of the reduced mitochondrial preparation indicated the presence of cytochrome P-450 in the preparation. 7. It concluded that the alpha-ecdysone C-20 hydroxylase system is a cytochrome P-450-deendent mono-oxygenase.

scholarly journals Maximum activities and properties of glucose 6-phosphatase in muscles from vertebrates and invertebrates

1981 ◽  
Vol 198 (3) ◽  
pp. 621-629 ◽  
Author(s):  
B Surholt ◽  
E A Newsholme
Keyword(s):  
Flight Muscle ◽  
Schistocerca Gregaria ◽  
Pectoral Muscle ◽  
Hydrolytic Activity ◽  
Strong Positive Correlation ◽  
Ph Optimum ◽  
Futile Cycle ◽  
Avian Muscle ◽  
Glucose Phosphorylation ◽  
Almost All

1. The maximum catalytic activities of glucose 6-phosphatase were measured in a large number of muscles from vertebrates and invertebrates. The activities range from less than 0.1 to 8.0 mumol/min per g fresh wt. at 30 degrees C: the highest activity, observed in the flight muscle of the wasp (Vespa vulgaris), is similar to that in rat liver. The hydrolytic activity was shown to be specific towards glucose 6-phosphate. 2. The pH optimum was 6.8 and the Km was approx. 0.6 mM (flight muscle of a moth). 3. Almost all of the glucose 6-phosphatase activity from extracts of the flight muscle of a moth and the pectoral muscle of a pigeon were recovered in the cytosolic fraction (i.e. 150,000 g supernatant). 4. During development of the locust (Schistocerca gregaria), the activity of the phosphatase in the flight muscle increased during the first 3 days after the final moult. 5. The activity of glucose 6-phosphatase from insect and avian muscle was separated from that of non-specific phosphatase on a Bio-Gel P-100 column. 6. For the activities from 63 muscles, there was a strong positive correlation between those of glucose 6-phosphatase and hexokinase, but no correlation between the activities of glucose 6-phosphatase and fructose bisphosphatase. It is suggested that the role of glucose 6-phosphate in muscle is either to produce glucose from glucose 6-phosphate derived from glycogen or to provide the enzymic basis for a substrate (‘futile’) cycle between glucose and glucose 6-phosphatase in muscle to improve the sensitivity of the mechanism that regulates the rate of glucose phosphorylation.

FERMENTATIVE HYDROLYTIC PREPARATION METHODS OF CEREAL WORT FOR ALCOHOL FERMENTATION

1970 ◽  
pp. 52-56
Author(s):  
Е. M. Serba ◽  
М. B. Overchenko ◽  
L. V. Rimareva ◽  
N. I. Ignatova ◽  
А. E. Orekhova ◽  
...  
Keyword(s):  
Alcoholic Fermentation ◽  
Raw Materials ◽  
Alcohol Concentration ◽  
Activity Level ◽  
Optimal Dosage ◽  
Main Role ◽  
Alcohol Fermentation ◽  
Ph Optimum ◽  
Amylolytic Enzyme ◽  
Enzyme Preparations

In the production of alcohol in the preparation of grain raw materials for fermentation, the main role is given to enzyme preparations of amylolytic action, which are key enzymes that catalyze the hydrolysis of starch. Amylolytic enzyme preparations with a different composition of enzymes and their level of activity, a mechanism of biocatalytic effect on starch, and a range of thermal and pH optimum are widely represented on the Russian market. The development of optimal conditions for the preparation of grain wort, the rational selection and dosage of concentrated enzyme preparations, the properties of which correspond to the parameters of the technological process, will ensure the effective preparation of starch for fermentation, and increase the profitability of alcohol production. The aim of this work was to study the influence of enzyme preparations of amylolytic action and the conditions of their use on the efficiency of the process of alcoholic fermentation and the yield of the final product, ethanol. The effect of various dosages of enzyme preparations of glucoamylase action, with a different ratio of the main enzyme glucoamylase and minor enzyme α-amylase, as well as methods for preparing wheat wort on the process of alcoholic fermentation, was studied. It was found that the enzyme preparation, the source of glucoamylase, in which α-amylase was present in a ratio of 15: 1 (in terms of activity level), turned out to be more effective in fermenting prepared wheat wort: its optimal dosage was 8 units. GLS / g starch. The presence of a sufficient amount of α-amylase in this preparation compensated for the dosage of thermostable α-amylase. The alcohol concentration in the mash was 10.2% vol., The alcohol yield was 67.9 cm3 / 100 g of starch. When glucoamylase with a lower ratio of the main and minor enzyme (75: 1) was used at the saccharification stage, an increase in the wort fermentation depth was observed with an increase in the concentration of glucoamylase to 9-10 units of GLS / g and α-amylase to 0.5 units. AC / g. It was also found that an increase in the duration of enzymatic-hydrolytic preparation of the wort had a positive effect on the fermentation process, the alcohol concentration in the mash increased to 10.2 vol.%. It was shown that the introduction of proteases into the wort helps to reduce the viscosity of grain wort, enriching it with assimilable yeast amino acids, which leads to an increase in the yield of alcohol. It has been confirmed that the synergy of the action of enzymes of amylolytic and proteolytic effects on polymers of grain raw materials allows to increase the efficiency of their conversion to ethanol. The conditions of enzymatic-hydrolytic processing of grain raw materials for fermentation are developed. The use of the digestion stage did not significantly affect the fermentation results of wheat wort.

THE HISTOLOGY OF THE GIANT FIBRE SYSTEM IN THE ABDOMINAL VENTRAL NERVE CORD OF THE DESERT LOCUST

1970 ◽  
Vol 102 (9) ◽  
pp. 1163-1168 ◽  
Author(s):  
W. D. Seabrook
Keyword(s):  
Single Cell ◽  
Nerve Cord ◽  
Ventral Nerve Cord ◽  
Schistocerca Gregaria ◽  
Desert Locust ◽  
Giant Fibre ◽  
Metathoracic Ganglion ◽  
Giant Fibres ◽  
Giant Fibre System

AbstractSchistocerca gregaria possess four neurones of giant fibre proportions within the abdominal ventral nerve cord. These fibres arise from single cell bodies in the terminal ganglionic mass and pass without interruption to the metathoracic ganglion. Fibres become reduced in diameter when passing through a ganglion. Branching of the giant fibres occurs in abdominal ganglia 6 and 7.

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