Acid phosphatase, inhibition

One of the most regrettable characteristics of enzyme inhibition-based biosensors is the different inhibition degree caused by different inhibitors. An example of this behavior is an amperometric acid phosphatase inhibition-based biosensor proposed by Mazzei et al., which shows different inhibition efficiency towards organophosphorus and carbamate compounds (the latter with a weaker inhibition efficacy and higher detection limits, as a result). 

Sample Collection and Enzyme Stability. Serum samples are collected with chemically clean, sterile glassware. Blood is allowed to clot at room temperature, the clot is gently separated from the test tube with an applicator stick, and the blood is centrifuged for 10 minutes at 1,000 g. If the red cells are known to contain the enzymes whose activity is being measured, as in the case of LD, even slightly hemolyzed serums must be discarded. When acid phosphatase is to be measured, the serum should be placed immediately in ice and processed as soon as possible, or it should be acidified by the addition of a small amount of sodium citrate. Anticoagulants such as EDTA, fluoride and oxalate inhibit some serum enzymes. However, heparin activates serum lipoprotein lipase. 

Red blood cells also contain sufficient acid phenylphospha-tase for mild hemolysis to cause false elevations. Therefore, inhibitors such as ethanol, formaldehyde, copper sulfate> and 1-tartrate have been used to inhibit selectively the enzyme of one or more tissues and enhance the specificity of the test (101). Ethanol is unsuitable because it inhibits the enzyme from erythrocytes and prostate simultaneously, and because it yields serum activities which correlate poorly with prostatic disease. Formaldehyde inhibits the erythrocytic enzyme and has been said to yield clinically satisfactory results. The copoper resistant acid phosphatase of serum is elevated by metastatic carcinoma of the breast, as well as by other metastatic cancers, and is also elevated by a wide variety of non-cancerous diseases. 

Figure 5 Model of phosphorus (P) deficiency-induced physiological changes associated with the release of P-mobilizing root exudates in cluster roots of white lupin. Solid lines indicate stimulation and dotted lines inhibition of biochemical reaction sequences or mclaholic pathways in response to P deliciency. For a detailed description see Sec. 4.1. Abbreviations SS = sucrose synthase FK = fructokinase PGM = phosphoglueomutase PEP = phosphoenol pyruvate PE PC = PEP-carboxylase MDH = malate dehydrogenase ME = malic enzyme CS = citrate synthase PDC = pyruvate decarboxylase ALDH — alcohol dehydrogenase E-4-P = erythrosc-4-phosphate DAMP = dihydraxyaceConephos-phate APase = acid phosphatase.

Associated with NA depletion from host nuclei was the diminution and deformation of these nuclei, significant depletion of host nucleoskeletal protein lamin b, and significant reductions in levels of host cell RNA, protein and acid phosphatase activity. In contrast, host infiltrating cells showed no such changes, supporting an infected cell-specific effect. These observations support the possibility that NA are synthesized by the parasite and modulate host nuclear functions. However, release of other parasite secretory products, not analysed, might have been inhibited in these experiments also. 

Abbreviations. a-M, a-mannosidase AP, acid phosphatase as-ni-ATPase, anion-stimulated, nitrate-inhibitable ATPase CCR, NAD(P)H-dependent cytochrome oreduc-tase cs-vi-ATPase, cation-stimulated, vanadate-inhibitable ATPase, CAT, catalase GS 1/11, glucan synthase 1 or 11 IDPase, inosine diphosphatase cs-PPase, cation-stimulated pyrophosphatase RNA polymerase, DNA-dependent RNA polymerase TP-25, 25 kDa tonoplast integral protein. 

It is not clear whether V(V) or V(IV) (or both) is the active insulin-mimetic redox state of vanadium. In the body, endogenous reducing agents such as glutathione and ascorbic acid may inhibit the oxidation of V(IV). The mechanism of action of insulin mimetics is unclear. Insulin receptors are membrane-spanning tyrosine-specific protein kinases activated by insulin on the extracellular side to catalyze intracellular protein tyrosine phosphorylation. Vanadates can act as phosphate analogs, and there is evidence for potent inhibition of phosphotyrosine phosphatases (526). Peroxovanadate complexes, for example, can induce autophosphorylation at tyrosine residues and inhibit the insulin-receptor-associated phosphotyrosine phosphatase, and these in turn activate insulin-receptor kinase. 

The cytotoxicities of okadaic acid as EC50 against the P388 and L1210 cell lines are 1.7 nano-molar and 17 nano-molar, respectively. Additionally, okadaic acid strongly inhibits protein serine/threonine phosphatase 1,2A,... 

AGIRE computer program for, 249, 79-81, 225-226 comparison to analysis based on rates, 249, 61-63 complex reactions, 249, 75-78 experimental design, 249, 84-85 inhibitor effects, 249, 71-75 potato acid phosphatase product inhibition, 249, 73-74 preliminary fitting, 249, 82-84 prephenate dehydratase product inhibition, 249, 72-73 product inhibition effects, 249, 72-73 prostate acid phosphatase phenyl phosphate hydrolysis, 249, 70 reactions with two substrates, 249, 75-77 reversible reactions, 249, 77-78 with simple Michaelian enzyme, 249, 63-71 [fitting equations, 249, 63] with slow-binding inhibitors, 249, 88 with unstable enzymes, for kinetic characterization, 249, 85-89. 

Administration of 1 and 3 mg Sn/kg body weight to rats resulted in inhibition of various enzymes, including hepatic succinate dehydrogenase and the acid phosphatase of the femoral epiphysis. Tin also appears to interact with the absorption and metabolism of biological essential metals such as copper, zinc, and iron and to influence heme metabolism. ... 

Enzymes activities are particularly sensitive to the anticoagulant used in collecting the specimen. Heparin inhibits acid phosphatase (W16) and muramidase (Z5). Amylase activity is inhibited by oxalate or citrate (MIO), and lactic dehydrogenase and acid phosphatase lose activity in oxalate (C2). Alkaline phosphatase is stable in oxalate, oxalate-fluoride, or heparin, but 25 mAf citrate inhibits 50% of the activity, and as little as 50 mlf EDTA is completely inhibitory (B19). Leucine aminopeptidase is inhibited by EDTA, as is creatine phosphokinase (F3). Amylase activity has been reported to be only 83% of that in serum when oxalate or citrate-plasma is used (MIO). Heparin plasma appears to have no inhibitory effect. Despite the fact that clotting factor V is not stable in oxalate or EDTA, these are often used as anticoagulants to obtain plasma for prothrombin determinations (Z2, Z4). 

K.H. Lau, T.K. Freeman, D.J. Baylink, A proposed mechanism of the mitogenic action of fluoride on bone cells. Inhibition of the activity of an osteoblastic acid phosphatase, Metabolism 38 (1989) 858-868. 

Acid phosphomonoesterase (EC 3.1.3.2). Milk contains an acid phosphatase which has a pH optimum at 4.0 and is very heat stable (LTLT pasteurization causes only 10-20% inactivation and 30 min at 88°C is required for full inactivation). Denaturation of acid phosphatase under UHT conditions follows first-order kinetics. When heated in milk at pH 6.7, the enzyme retains significant activity following HTST pasteurization but does not survive in-bottle sterilization or UHT treatment. The enzyme is not activated by Mg2+ (as is alkaline phosphatase), but it is slightly activated by Mn2+ and is very effectively inhibited by fluoride. The level of acid phosphatase activity in milk is only about 2% that of alkaline phosphatase activity reaches a sharp maximum 5-6 days post-partum, then decreases and remains at a low level to the end of lactation. 

T15. Clinical Application of Differential Enzyme Inhibition Human blood serum contains a class of enzymes known as acid phosphatases, which hydrolyze biological phosphate esters under slightly acidic conditions (pH 5.0) ... 

Acid phosphatases are produced by erythrocytes, the liver, kidney, spleen, and prostate gland. The enzyme of the prostate gland is clinically important, because its increased activity in the blood can be an indication of prostate cancer. The phosphatase from the prostate gland is strongly inhibited by tartrate ion, but acid phosphatases from other tissues are not. How can this information be used to develop a specific procedure for measuring the activity of the acid phosphatase of the prostate gland in human blood serum ... 

Beckman et al. (28) have studied the electrophoretic separation of the acid phosphatase activity in tissue extracts on starch gel at pH 8. They described four electrophoretic bands A, B, C, and D. Table IV (28) shows the distribution of activity in different organ extracts. The ABD pattern predominated in kidney BD in liver, intestine, heart, and skeletal muscle B in skin and D in pancreas. The C component was present in a large number of placentae but not in other adult organs. All four electrophoretic components were inhibited by d-(- -)-tartrate A contained sialic acid, D had a lower pH optimum and was more heat resistant than A, B, and C. Components C and D showed parallel electrophoretic behavior. In human skin fibroblasts grown in tissue culture, the acid phosphatase was generally high and the most common pattern was BD. Almost every culture showed some activity. The BD... 

Most investigators utilize p-nitrophenyl or a-naphthyl phosphate as substrate. The determination of serum prostatic acid phosphatase was developed by Fishman and Lemer (34) based on the d-(+)-tartrate inhibition of prostatic enzyme discussed below. Babson et al. (35, 36) demonstrated that a-naphthyl phosphate was much more easily split by prostatic than red cell phosphatase. Table V (35) shows the results obtained when prostatic or red cell phosphatase was added to human serum which had been incubated at pH 8.6 for 1 hr at 37° to destroy all endogeneous phosphatase activity. The table shows the superiority of a-naphthyl phosphate as substrate. 

Reiner and his colleagues (40) demonstrated that fluoride inhibition of prostatic acid phosphatase showed interesting and unexpectedly complex kinetics. The unusual nature of the inhibition can readily be appreciated from Fig. 4 (40). As the fluoride concentration is increased over a 1000-fold range, the extent of inhibition rises and then subsequently falls with a further increase of inhibitor. At lower fluoride concentration, the inhibition is clearly competitive. Two possibilities were explored for an explanation of these unusual concentration effects of inhibition. There could be two forms of fluoride in the reaction mixtures the inhibitory form and the second which predominates at higher... 

Vescia and Chance Hi) demonstrated that fluoride and tartrate inhibition vide infra) of acid phosphatase showed completely different kinetics when the hydrolysis of phenyl phosphate was compared with transphosphorylation from this substrate to glucose. Figures 5 and 6 (41) show that fluoride inhibition is competitive when the data are plotted according to Lineweaver and Burk. However, the inhibition is noncompetitive with respect to transphosphorylation of the same substrate to glucose. The authors suggested that there are two distinct sites... 

Inhibition of Prostatic Acid Phosphatase by Various Hydroxycarboxylic... 

Fia. 7. Inhibition of prostatic acid phosphatase by D-(+)-tartaric acid. The reaction mixtures all contained equivalent amounts of the enzyme preparation, the indicated concentration of substrate (pH 5.0), 0.05 iff acetate buffer (pH 5.0), and tartaric acid (pH 5.0) total volume, 4.5 ml. Each point represents average values of determinations made with 5 X 10-5 iff and 10 X 10"5 iff tartaric acid except in the case of /3-glycerophosphate for which 1 X 10 iff and 2 X 10 iff tartaric acid was used. From Kilsheimer and Axelrod (4 ). 

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