Purification and some properties of an atypical alkaline p-nitrophenylphosphate phosphatase activity from Halobacterium halobium

• 1.1. An alkaline p-nitrophenylphosphate phosphatase has been purified 440-fold from extracts of Hatobacterium halobium.
• 2.2. The enzyme has an apparent molecular weight of 24,000.
• 3.3. A K_m value for p-nitrophenylphosphate of 1.12mM has been found under optimal conditions.
• 4.4. The enzyme is selectively activated and stabilized by Mn²⁺.
• 5.5. It requires high salt concentrations for stability and maximum activity.
• 6.6. It displays an unusual restricted substrate specificity of 25 phosphate esters tested, only phosphotyrosine and casein were hydrolysed besides p-nitrophenylphosphate.

     

A comparative study on the biological properties of venoms from juvenile and adult common tiger snake (Notechis scutatus) venoms

• 1.1. The biological properties of venoms from juvenile and adult common tiger snakes (Notechis scutatus) were compared.
• 2.2. The lethality, procoagulant activity and enzymatic activities of the juvenile venom were not substantially different from those of the adult venom.
• 3.3. Electrophoretic studies, however, indicated some minor differences in the protein composition of the juvenile and adult venoms.

     

Hemorrhagic and mojave toxins in the venoms of the offspring of two mojave rattlesnakes (Crotalus scutulatus scutulatus)

• 1.1. The venoms of two Mojave rattlesnakes and those of their offsprings were analyzed for Mojave toxin and hemorrhagic toxin.
• 2.2. The venom of one female, collected in Pima County, Arizona, and the venoms of her six offspring contained hemorrhagic toxin but not Mojave toxin (venom B).
• 3.3. The venom of the second female, captured in El Paso County, Texas, contained both toxins (A + B venom). Of her 10 offspring, five contained venom with both toxins, two had hemorrhagic toxin only, and three contained neither toxin.
• 4.4. Venoms that caused hemorrhage also inactivated complement. A pool of the venoms of the venom B offspring was less toxic than adult pooled venom A.

     

Alkaline phosphatase from Basidiobolus haptosporosus: Comparison with alkaline phosphatases from rainbow lizard and man

• 1.1. DEAE-cellulose chromatography of mycelial alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) from Basidiobolus haptosporosus, produced three iso-enzymes “A”, “B” and “C”.
• 2.2. Fraction “A” was further characterized and showed maximum activity at pH 10 in 0.1 M sodium carbonate-bicarbonate buffer.
• 3.3. The enzyme was stimulated by Mg²⁺, Co²⁺ and Mn²⁺ and inactivated by Zn²⁺, Cu²⁺, EDTA, citrate and tartrate.
• 4.4. Phosphate ions inhibited it competitively, phenylalanine uncompetitively and urea noncompetitively.
• 5.5. It was heat stable for 60 min at 37°C but labile above 55°C.
• 6.6. Its K_m with p-nitrophenylphosphate was 0.5 mM; its estimated molecular weight was 160,000.
• 7.7. The results are compared with the properties of alkaline phosphatases from the rainbow lizard and man and discussed in terms of a triadic association between the fungus, the lizard and man.

     

Seasonal changes of water,electrolytes and aldosterone levels in blood serum,brain and kidney of the Egyptian cobra “Naja haje haje”

 

• 1.The levels of water, Na, K, Ca and Mg in blood serum, brain and kidney and aldosterone level in blood of Naja haje haje were studied during the different phases of the annual cycle.
• 2.The water content in the tissues studied displayed only minor changes as the animals passed from one phase to the other.
• 3.A significant increase in Na was recorded in the brain during the different phases indicating a depressed sodium pump, whereas the blood Na level showed a significant decrease during hibernation.
• 4.K increased in blood serum, brain and kidney during hibernation, while a nonsignificant decrease was found in blood serum during arousal. The brain may act as a potassium reservoir.
• 5.An increase in Ca and Mg concentration was recorded in blood serum, brain and kidney during prehibernation and hibernation. The data suggested a homeostatic function in the transport and metabolism of these cations.
• 6.Aldosterone exhibited a highly significant decrease especially during hibernation. The aldosterone regulation of ionic composition is discussed.
• 7.Na/K and Ca/Mg ratios in the brain may explain the decreased excitability during winter torpor.

     

Lipid changes in blood serum and tissues of the Egyptian Cobra “Naja haje haje” during the hibernation cycle

• 1.Total lipids, free fatty acids, triglycerides, phospholipids and total cholesterol in blood serum, liver, brain, cardiac and skeletal muscles of Naja haje haje were determined during the different phases of the hibernation cycle.
• 2.A sharp decrease in the level of total lipids of blood serum and all tissues occurred during hibernation. Upon arousal, lipogenesis is commonly restored.
• 3.Elevated concentrations of serum free fatty acids predominated in pre-hibernation and hibernation periods, while the tissues recorded highly significant declines during hibernation.
• 4.Occurrence of marked decreases in triglycerides contents of serum and tissues except the cardiac muscles in the hibernation and arousal phases.
• 5.Sharp increases in the phospholipid contents of blood and the selected tissues were recorded during hibernation. The level declined in both liver and cardiac muscles in arousing animals.
• 6.Total cholesterol level was lowered in blood during hibernation. The cardiac muscles showed a highly significant decrease while liver, brain and skeletal muscles showed elevations in the same phase.

     

Properties of an alkaline phosphatase from the dinoflagellate Peridinium cinctum

• 1.1. Alkaline phosphatase (EC 3.1.3.1) from the dinoflagellate Peridinium cinctum, the Lake Kinneret bloom alga, has been partially purified by gel filtration.
• 2.2. The enzyme could be easily extracted using a distilled water/chloroform mixture suggesting that the alkaline phosphatase of Peridinium is particularly labile.
• 3.3. The molecular weight of the enzyme was estimated as 158,000 ± 5000. The enzyme showed a broad pH optimum (in the range pH 8.0–8.5), had a K_m of 0.45 mM for p-nitrophenylphosphate as substrate and was stable to repeated freeze/thawing cycles.
• 4.4. The enzyme was strongly activated by Mg²⁺ whereas Zn²⁺ (and to a lesser extent Cd²⁺) was an effective inhibitor of the enzyme. Cu²⁺ activated the enzyme at low concentrations, although at higher concentrations inhibited the enzyme. This effect of metals on the Peridinium alkaline phosphatase could be environmentally important since underwater hot springs, containing high concentrations of copper, enter the lake.

     

Two acid phosphatases in sea urchin eggs and embryos

• 1.1. Two types of acid phosphatases from sea urchin eggs and embryos were studied in three Japanese species, Hemicentrotus pulcherrimus, Anthocidaris crassispina and Pseudocentrotus depressus.
• 2.2. Acid phosphatase type 1, designated AcP-1, hydrolysed only flavin mononucleotide besides p-nitrophenylphosphate. The activity of AcP-1 was not inhibited by NaF and tartrate. This enzyme showed molecular weight between 14,000 and 16,000 by gel filtration through Sephadex G-75.
• 3.3. The higher molecular weight type of acid phosphatase, designated AcP-2, showed relatively high substrate specificity toward ADP and ATP. Molecular weight of AcP-2 ranged from 42,000 to 48,000 by gel filtration through Sephadex G-100.
• 4.4. Some properties of AcP-2 from Sphaerechinus granularis embryos are also described.

     

Purification and some properties of a Mg2+-activated acid phosphatase from rat testis

• 1.1. The acid phosphatase (AcPase, EC 3.1.3.2) IV from rat testicular tissue was purified to apparent homogeneity.
• 2.2. The enzyme displays a native molecular weight of 70 kDa determined on gel permeation chromatography on a Sephadex G-100 column and 68 kDa using linear 5–20% sucrose density gradient centrifugation. The subunit molecular weight on SDS-PAGE analysis is 67 kDa, suggesting that the enzyme is a monomeric protein.
• 3.3. The enzyme does not bind to Concanavaline A-Sepharose 4B column, indicating that it is not a glycoprotein.
• 4.4. The rat testis AcPase IV is a metal activated enzyme in which Mg²⁺ is the metal activating agent with a K_a, = 0.88 × 10⁻³ M. The Michaelis constant for p-nitrophenylphosphate, in the presence of saturating concentrations of Mg²⁺ ions, is 0.23 × 10⁻³ M.
• 5.5. The enzyme preferentially hydrolizes p-nitrophenylphosphate, phenylphosphate and ATP.

     

Evidence concerning a possible steady state rate equation for E. coli alkaline phosphatase

• 1.1. Steady state data was obtained for alkaline phosphatase over a wide range of experimental conditions using two substrates, four inhibitors, two modifiers and several pH, ionic strength and temperatures values.
• 2.2. The data was fitted by rational functions of degree 1:1, 2:2 and 3:3 using a non-linear regression program and then the F-test was used to assess the goodness of fit.
• 3.3. A proportion of the curves could only be fitted by 2:2 functions but many of them could be adequately fitted by 1:1 functions.
• 4.4. No statistically significant improvement in fit occurred with 3:3 functions.
• 5.5. Data was simulated using a computer program to see what sort of curves could be generated by a two sites mechanism proposed for alkaline phosphatase and this study showed that it is difficult to detect cubic terms in this rate equation.
• 6.6. It was concluded that alkaline phosphatase does not obey Michaelis-Menten kinetics. Rather, the steady state data require a mechanism of at least second degree but do not exclude a rate equation of third degree.

     

The action of the venom of Philanthus triangulum F. on an insect visceral muscle

• 1.1. Low concentrations (0.05−0.38 BU/ml) of a crude venom extract from P. triangulum F. potentiate nerve-evoked contractions of the locust hindgut, possibly due to contamination of the venom preparation with proctolin.
• 2.2. Higher venom concentrations inhibit nerve-evoked contractions to a dose-independent plateau level.
• 3.3. The venom has no effect on responses to bath-applied proctolin, but responses to bath-applied L-glutamate are inhibited.
• 4.4. Spontaneous contractions are unaffected by the venom.
• 5.5. It is concluded that the plateau contractions are the result of excitation by non-glutamatergic transmission, and are possibly the result of proctolin release.

     

Prostaglandin E2/parathyroid hormone-induced suppression of alkaline phosphatase activity is mediated by protein kinase C

• 1.1. Bone resorptive factors, prostaglandin E2 and parathyroid hormone are shown to suppress alkaline phosphatase activity in a rat osteoblastic cell line.
• 2.2. Phorbol myristate acetate, but not dibutyryl cAMP or calcium ionophore can suppress alkaline phosphatase activity.
• 3.3. The protein kinase C inhibitors (H89, staurosporine) are able to block the suppression of alkaline phosphatase activity induced by prostaglandin E2 and parathyroid hormone.
• 4.4. These data suggest that protein kinase C is involved in the inhibition of alkaline phosphatase activity induced by prostaglandin E2 and parathyroid hormone.

     

Isolation of a hemorrhagic toxin from the venom of Agkistrodon contortrix laticinctus (broad-banded copperhead) and pathogenesis of the hemorrhage induced by the toxin in mice

• 1.1. A hemorrhagic toxin was isolated from the venom of Agkistrodon contortrix laticinctus (Broad-Banded Copperhead) by Sephacryl S-200 HR column chromatography followed by high performance chromatography on Waters DEAE 5PW and protein Pak 125 columns.
• 2.2. Homogeneity was determined by the presence of a single band in acrylamide gel electrophoresis with silver staining.
• 3.3. ACL hemorrhagic toxin I has a molecular weight of about 29,000, is slightly acidic, and is a metalloprotease with activity towards the substrates N,N-dimethylcasein and bovine fibrinogen. Although the toxin is able to hydrolyze fibrinogen in vitro, it does not possess any defibrinogenating activity in vivo whereas the crude venom does show this activity. It has similar cleavage specificities to other snake venom hemorrhagic toxins.
• 4.4. ACL hemorrhagic toxin I causes hemorrhage of rapid onset, present within 5 min of intramuscular injection into mice, and the pathogenesis is one of hemorrhage per rhexis in which capillary endothelial cells are ruptured.

     

Correlation of acid phosphatase but not alkaline phosphatase activity with myelination in the developing mouse brain

• 1.1. Neonatal mice received subcutaneous injections of buffer, thiourea (TU) or propylthiouracil (PTU).
• 2.2. The PTU-treated mice were sacrificed on postnatal day 14 (P14) and the TU-treated mice on P28.
• 3.3. Brain weights of the TU- and PTU-treated mice were not significantly different from the controls.
• 4.4. Acid but not alkaline phosphatase activity in the braistem decreased after TU and PTU treatment.
• 5.5. Myelination as indicated by intensity of luxol fast blue staining was weaker in drug-treated animals.
• 6.6. The level of myelin marker enzyme, 2′,3′-cyclic nucleotide 3′-phosphohydrolase, was lower in the brainstem of PTU-treated animals.
• 7.7. The results suggest a correlation between acid phosphatase but not alkaline phosphatase activity with myelination in the developing mouse brain.

     

Kinetic studies of the transphosphorylation reactions catalyzed by alkaline phosphatase from E. coli: Hydrolysis of p-nitrophenyl phosphate and o-carboxyphenyl phosphate in presence of tris

• 1.1. Transphosphorylation of p-nitrophenyl phosphate and o-carboxyphenyl phosphate to Tris, has been studied at alkaline and acid pH.
• 2.2. The rate of release for all reactions products was Tris-dependent for both substrates, with a slight maximum for phenol at alkaline pH. These dependences have been analyzed from a mechanistic standpoint.
• 3.3. Individual constants of rate of a simple transphosphorylation mechanism have been determined.
• 4.4. At high Tris concentrations (> 1.0 M) a slight competitive inhibition has been observed.
• 5.5. Inhibition in NH₄⁺-NH₃Cl buffer has been found at alkaline pH but not at acid pH. It would therefore seem that the non-protonated NH₂ group of Tris is responsible for inhibition.
• 6.6. The results suggest the formation of complexes between Tris and the enzyme. Other possible alternatives are also analyzed.

     

Alteration of the protein composition of bothrops jararaca venom and venom gland by isoproterenol treatment

• 1.1. The protein composition of Bothrops jararaca venom and venom gland was analyzed through SDS-PAGE, after isoproterenol (IPR) treatment.
• 2.2. Some proteins (47, 48, 57 and 72 kDa) were detected in the gland homogenate from the control but not from the IPR-treated samples.
• 3.3. Three proteins (26.5, 44.5 and 53 kDa) were detected in the venom gland from IPR-treated snakes but not from the venom gland from the control.
• 4.4. In the venom samples proteins of 41 and 74 kDa were detected only in the IPR treated samples, while proteins of 17 and 28 kDa were detected only in the control.
• 5.5. The biological activity of the venom did not change with IPR treatment.

     

Effects of short chain fatty acid,sodium butyrate,on osteoblastic cells and osteoclastic cells

• 1.1. Sodium butyrate increased alkaline phosphatase (ALP) activity of cloned osteoblastic cell line MC3T3-El by the stimulation of de novo enzyme synthesis.
• 2.2. Sodium butyrate did not affect mature osteoblastic cells but affected preosteoblastic cells.
• 3.3. Sodium butyrate decreased tartrate resistant acid phosphatase (TRACP)-positive multinucleated cells (MNC) formation from bone marrow cells. This related to the cytotoxicity of sodium butyrate on bone marrow cells.

     

Serum chemistry profiles for Lechwe waterbucks (Kobus leche): Variations with age and sex

• 1.1. Over an 8-year period, 19 biochemical parameters have been determined at various ages in the blood serum of 92 clinically healthy Lechwe waterbucks (Kobus leche), 33 males and 59 females.
• 2.2. Significant differences have been noted with age. In neonates, the lowest values of total proteins, glucose, creatinine, urea, AST, ALT and iron have been noted; the highest ones have been seen for cholesterol, alkaline phosphatase, calcium and phosphorus.
• 3.3. With regard to sex, raised values of glucose, urea, alkaline phosphatase and ALT, and lowered values of cholesterol, have been noted in juvenile females compared with males of the same age.
• 4.4. In adult females, higher levels of urea and cholesterol and lower levels of glucose, triglycerides and natrium have been recorded compared with males.
• 5.5. With sex and age, no significant changes have been found in the levels of GGT, magnesium, chlorides and copper.
• 6.6. Out findings are discussed with those abstracted from the literature for related species.

     

The haemocytes and the activities of leucine aminopeptidase and alkaline phosphatase during development of Ceratitis capitata: Specific secretion of a leucine aminopeptidase isozyme

• 1.1. The types of haemocytes during larval development were studied.
• 2.2. The developmental profile of leucine aminopeptidase and alkaline phosphatase was studied. The maximum LAP activity was found to be in early larval development, while the maximum alkaline phosphatase during the white pupal stage.
• 3.3. These activities were compared with those determined in cell-free haemolymph.
• 4.4. Both hydrolytic enzymes have been found histochemically in the prohaemocytes and in the plasmatocytes.
• 5.5. In cultured haemocytes experiments it was found that 64% of the total LAP activity was secreted into the incubation medium, while electrophoretic analysis of released LAP activity demonstrated that only LAP A isozyme was secreted.
• 6.6. Based on the above results we suggest that both hydrolytic enzymes are functionally important throughout larval development.

     

Phosphorylcholine as a unique substrate for human intestinal alkaline phosphatase

• 1.1. The enzymatic nature of human liver, bone, placental and intestinal alkaline phosphatases (ALPs) were investigated with phosphorylcholine (PC), phosphoryl-ethanolamine, pyridoxal-5'-phosphate and p-nitrophenylphosphate at a weakly alkaline pH.
• 2.2. The apparent K_m value of the intestinal ALP with PC was the highest of all ALPs tested. Intestinal ALP hydrolyzes PC the most and has higher affinity for choline as a transphorsphorylating acceptor than the other ALPs. In addition, the intestinal ALP activity with PC was most susceptible to Na₂HPO₄, in the tested ALPs.
• 3.3. The present results suggest that PC is a unique substrate for human intestinal ALP, which may be related to the metabolism of PC or choline as part of phosphatidylcholine.