Interplay between acid phosphatase and cysteine proteases in mediating vitellin degradation during early embryogenesis of Periplaneta americana

The influence of L-carnitine in vitro on the lysosomal cysteine proteinase activity and stability of the lysosomal membrane of the liver homogenates of intact sexually Mature female rats of Wistar line weighing 280-330 g were studied. In the experimental groups isolated lysosomes were incubated in vitro in a solution of L-carnitine during 1, 2 and 4 hours, in the control groups in vitro incubation was carried out in a medium of isolating solution. The activity of ca-thepsins B, L and H was investigated by spectrofluorimetric method of Barrett & Kirschke in two fractions - lysosomal and outside of lysosomes. The activity of acid phosphatase was used as the main marker of a membrane labilization. In vitro incubation of lysosomes showed that carnitine at a concentration of 5 mM increases the total activity of cathepsin B in a one-hour incubation at 73,2% (p=0,008), cathepsin L in a two- and four-hour incubation - at 77,7% (p=0,005) and 42,3% (p=0,013) respectively, and reduces the overall activity of the cathepsin H in a one-hour incubation at 200,0% (p=0,008), in a two-hour - by 67,9% (p=0,05), in a four-hour -27,1% (p=0,02). In addition, incubation in 5 mM L-carnitine solution leads to an increase of unsedimentable activity and fall sedimentaries activity for cathepsin L in a two-hour, and for acid phosphatase - in a two - and four-hour exposure. 5 mM L-carnitine in one - and two-hour incubation stabilizes lysosomal membrane (whereas increase in incubation time up to 4 hours leads to its damage) and increases the selective permeability of the lysosomal membrane for the studied cathepsins, to the greatest extent - for cathepsin H.

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Effect of L-arginine and L-name on lysosomal cysteine proteases activity and lysosomal membranes permeability in rat aorta

Kazan medical journal ◽

10.17750/kmj2016-250 ◽

2016 ◽

Vol 97 (2) ◽

pp. 250-255 ◽

Cited By ~ 1

Author(s):

A I Arapova ◽

M A Fomina

Keyword(s):

Acid Phosphatase ◽

Cathepsin B ◽

Cysteine Proteases ◽

Total Activity ◽

Rat Aorta ◽

Lysosomal Fraction ◽

Lysosomal Membranes ◽

Cathepsin H ◽

Male Wistar Rats ◽

Combined Drugs

Aim. To study the effect of L-arginine and its analogue N-nitro-L-arginine methyl ester (L-NAME) alone and in combination on lysosomal cysteine proteolysis and lysosomal membranes state in rat aorta.Methods. The study was performed on male Wistar rats kept under standard vivarium conditions and divided into three control and three experimental groups of 8 animals each. The experimental samples included groups with L-arginine and/or L-NAME administration. The indicators were studied in the rat aorta homogenate precipitating and non precipitating fractions. Acid phosphatase activity was determined by a standardized method of «end point», the cathepsins B, L and H activity was studied by spectrofluorimetric method.Results. When simulating the changes of nitric oxide synthesis level using L-arginine, the increase of the total cathepsins activity was detected, acid phosphatase lability coefficient was reduced, what is characterized by general lysosomal membranes stabilization. L-NAME group, in contrast, is characterized by a decrease in the cathepsin B and H activity indicators, differences from arginine group were observed in the cathepsin H in lysosomal and general fractions, lysosomal membrane is labile. Combined drugs administration reduces the total cathepsins activity, while there is an increase of the acid phosphatase total activity, all indicators suggest lysosomal membranes labilization.Conclusion. L-arginine at a dose of 500 mg/kg causes increase in the total cathepsins B, L and H activity in rat aorta due to lysosomal fraction; L-arginine action leads to lysosomal membranes stabilization; L-NAME group in cathepsin H shows a decrease in the cathepsins secretion level with decreased total activity due to both factions; combined administration of arginine + L-NAME group in cathepsin B is characterized by an increase in secretion due to lysosomes membrane labilization.

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Early and late endosomal compartments of Entamoeba histolytica are enriched in cysteine proteases, acid phosphatase and several Ras-related Rab GTPases

Molecular and Biochemical Parasitology ◽

10.1016/s0166-6851(99)00133-4 ◽

1999 ◽

Vol 103 (2) ◽

pp. 225-241 ◽

Cited By ~ 43

Author(s):

Lesly A Temesvari ◽

Edward N Harris ◽

Samuel L Stanley ◽

James A Cardelli

Keyword(s):

Acid Phosphatase ◽

Entamoeba Histolytica ◽

Cysteine Proteases ◽

Rab Gtpases

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Effect of L-arginine and carnitine on cathepsin L and H activity and lysosomal membranes permeability in myocardium in expressed hyperhomocysteinemia

Kazan medical journal ◽

10.17750/kmj2015-819 ◽

2015 ◽

Vol 96 (5) ◽

pp. 819-824

Author(s):

A S Il’icheva ◽

M A Fomina

Keyword(s):

Acid Phosphatase ◽

Cathepsin L ◽

Cysteine Proteases ◽

Total Activity ◽

Lysosomal Membranes ◽

Cathepsin H ◽

Spectrofluorimetric Method ◽

Male Wistar Rats ◽

Reduced Activity ◽

Positive Effect

Aim. To study the activity of lysosomal cysteine proteases (cathepsins L, H) and acid phosphatase, changing of permeability, stability of myocardial lysosomal membranes in rats in experimental expressed hyperhomocysteinemia model, and while administering L-arginine and carnitine. Methods. The study was performed on male Wistar rats kept on standard vivarium conditions divided into three control and three experimental groups of 8 animals each. Experimental samples were administered methionine, or combination of L-arginine and carnitine with methionine. The level of serum homocysteine was measured by ELISA. Cathepsin L and H activity was detected by spectrofluorimetric method. Acid phosphatase activity was recorded using the «end point» method. Results. In the model of expressed hyperhomocysteinemia the increase of cathepsin H total activity due to both lysosomal and nonlysosomal fractions was found. These changes were observed along with the general increase of lysosomal membranes permeability. When correcting hyperhomocysteinemia with L-arginine and carnitine a decrease of cathepsin L and H levels was noted as well as positive effect on the myocardial lysosomal membranes stability. Conclusion. Expressed hyperhomocysteinemia is accompanied by statistically significant increase of both lysosomal and cytoplasmic fractions of the cathepsin H activity, indicating the lysosomal membranes permeabilisation phenomenon; L-carnitine and arginine correct hyperhomocysteinaemia effects, leading to cathepsin L and H reduced activity and having a stabilizing effect on the lysosomal membranes of cardiomyocytes.

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HYDROLYZING ENZYME ACTIVITY IN THE BRAIN AND THE RETROCEREBRAL SYSTEM OF THE AMERICAN COCKROACH (PERIPLANETA AMERICANA (L)) WITH SPECIAL REFERENCE TO THE NEUROSECRETORY MATERIALS

Canadian Journal of Zoology ◽

10.1139/z64-027 ◽

1964 ◽

Vol 42 (2) ◽

pp. 281-293 ◽

Cited By ~ 2

Author(s):

Y. Y. Shao ◽

S. E. Dixon

Keyword(s):

Enzyme Activity ◽

Acid Phosphatase ◽

Phosphatase Activity ◽

Acid Phosphatase Activity ◽

Periplaneta Americana ◽

American Cockroach ◽

Soluble Form ◽

Neurosecretory System ◽

Acid Phosphatases ◽

The Brain

Some hydrolyzing enzymes have been investigated by histochemical and biochemical methods. Almost all neurons in the brain contain acid phosphatases. About half of the total activity is associated with subcellular particles. Most of the acid-phosphatase activity in the corpora cardiaca (CC) is associated with the neurosecretory materials (NSM). Elementary neurosecretory granules (ENG) have been isolated from the CC by fractionation and subfractionation procedures and subjected to in vitro enzyme activity studies. Electron micrographs of the subfraction having the highest acid-phosphatase activity are discussed in relation to three types or "stages" of ENG. About 80% of the total esterase activity in the brain and the retrocerebral system has been found to be in a "soluble" form. The remaining 20%, the "insoluble" forms, may be associated with subcellular organelles. The methods of Gomori and Burstone for acid-phosphatase localizations are compared, and differing results discussed. The possible roles or functions of acid phosphatases and esterases in the neurosecretory system are discussed.

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Erythrophagocytosis in the Liver in Hemolytic Anemias

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100664 ◽

1968 ◽

Vol 26 ◽

pp. 184-185

Author(s):

O. T. Minick ◽

E. Orfei ◽

F. Volini ◽

G. Kent

Keyword(s):

Acid Phosphatase ◽

Oxidative Damage ◽

Phosphatase Activity ◽

Kupffer Cells ◽

Acid Phosphatase Activity ◽

Common Type ◽

Red Cell ◽

Cell Fragments ◽

Reticulo Endothelial System ◽

Important Site

Hemolytic anemias were produced in rats by administering phenylhydrazine or anti-erythrocytic (rooster) serum, the latter having agglutinin and hemolysin titers exceeding 1:1000.Following administration of phenylhydrazine, the erythrocytes undergo oxidative damage and are removed from the circulation by the cells of the reticulo-endothelial system, predominantly by the spleen. With increasing dosage or if animals are splenectomized, the Kupffer cells become an important site of sequestration and are greatly hypertrophied. Whole red cells are the most common type engulfed; they are broken down in digestive vacuoles, as shown by the presence of acid phosphatase activity (Fig. 1). Heinz body material and membranes persist longer than native hemoglobin. With larger doses of phenylhydrazine, erythrocytes undergo intravascular fragmentation, and the particles phagocytized are now mainly red cell fragments of varying sizes (Fig. 2).

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The Demonstration of Human Prostatic Acid Phosphatase (Pap) With Phosphorylcholine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100090427 ◽

1976 ◽

Vol 34 ◽

pp. 88-89

Author(s):

José A. Serrano ◽

Hannah L. Wasserkrug ◽

Anna A. Serrano ◽

Arnold M. Seligman

Keyword(s):

Acid Phosphatase ◽

Prostate Gland ◽

Prostatic Acid Phosphatase ◽

Human Prostate ◽

Rat Tissues ◽

Specific Substrate ◽

Acid Phosphatases ◽

Lysosomal Acid ◽

Cacodylate Buffer

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.

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Prostatic Acid Phosphatase (PAP) Differentiated Ultracytochemically from Lysosomal Acid Phosphate in the Rat with a PAP/Specific Substrate, Phosphorylcholine

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115870 ◽

1975 ◽

Vol 33 ◽

pp. 450-451

Author(s):

W. Allen Shannon ◽

José A. Serrano ◽

Hannah L. Wasserkrug ◽

Anna A. Serrano ◽

Arnold M. Seligman

Keyword(s):

Acid Phosphatase ◽

Phosphate Buffer ◽

Prostatic Carcinoma ◽

Prostatic Acid Phosphatase ◽

Chemotherapeutic Agents ◽

Specific Substrate ◽

Acid Phosphate ◽

Lysosomal Acid ◽

Design And Synthesis ◽

New Chemotherapeutic Agents

During the design and synthesis of new chemotherapeutic agents for prostatic carcinoma based on phosphorylated agents which might be enzyme-activated to cytotoxicity, phosphorylcholine, [(CH3)3+NCH2CH2OPO3Ca]Cl-, has been indicated to be a very specific substrate for prostatic acid phosphatase (PAP). This phenomenon has led to the development of specific histochemical and ultracytochemical methods for PAP using modifications of the Gomori lead method for acid phosphatase. Comparative histochemical results in prostate and kidney of the rat have been published earlier with phosphorylcholine (PC) and β-glycerophosphate (βGP). We now report the ultracytochemical results.Minced tissues were fixed in 3% glutaraldehyde-0.1 M phosphate buffered (pH 7.4) for 1.5 hr and rinsed overnight in several changes of 0.05 M phosphate buffer (pH 7.0) containing 7.5% sucrose. Tissues were incubated 30 min to 2 hr in Gomori acid phosphatase medium (2) containing 0.1 M substrate, either PC or βGP.

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