LINGCOD MUSCLE PHOSPHOMONOESTERASES: I. ACID PHOSPHATASES THAT HYDROLYZEp-NITROPHENYL PHOSPHATE

1960 ◽  
Vol 38 (6) ◽  
pp. 605-612 ◽  
Author(s):  
Neil Tomlinson ◽  
R. A. J. Warren
Keyword(s):  
Metal Ions ◽  
The Other ◽  
Acid Phosphatases ◽  
Free Base ◽  
Ph Optimum ◽  
Ophiodon Elongatus ◽  
Nitrophenyl Phosphate ◽  
Diethylaminoethyl Cellulose ◽  
Ph Range ◽  
Acid Phosphomonoesterase

Five fractions (A to E), each possessing acid phosphomonoesterase activity, were separated from an aqueous extract of the muscle of lingcod (Ophiodon elongatus) by stepwise chromatography on diethylaminoethyl cellulose in the free-base form.Fraction A required Zn++or Mn++for activity, was inhibited by heparin, and had its pH optimum at 6.0. Fraction E required Zn++for activity, was not inhibited by heparin, and had its pH optimum at 5.5. Fractions B, C, and D did not require metal ions for activity, and were distinguished from each other by differences in response to pH, cysteine, ethylenediaminetetraacetate, fluoride, and tartrate.The pH range over which fraction A was active was shifted to slightly higher values when Mn++was the activator rather than Zn++. Also, A was inhibited strongly by cysteine when activated by Zn++, but not when activated by Mn++. Data are presented that indicate these differences were due to different properties of the activating ions, rather than to the presence in fraction A of two enzymes, one activated by Zn++and the other by Mn++.

LINGCOD MUSCLE PHOSPHOMONOESTERASES: I. ACID PHOSPHATASES THAT HYDROLYZEp-NITROPHENYL PHOSPHATE

1960 ◽  
Vol 38 (1) ◽  
pp. 605-612 ◽  
Author(s):  
Neil Tomlinson ◽  
R. A. J. Warren
Keyword(s):  
Metal Ions ◽  
The Other ◽  
Acid Phosphatases ◽  
Free Base ◽  
Ph Optimum ◽  
Ophiodon Elongatus ◽  
Nitrophenyl Phosphate ◽  
Diethylaminoethyl Cellulose ◽  
Ph Range ◽  
Acid Phosphomonoesterase

Five fractions (A to E), each possessing acid phosphomonoesterase activity, were separated from an aqueous extract of the muscle of lingcod (Ophiodon elongatus) by stepwise chromatography on diethylaminoethyl cellulose in the free-base form.Fraction A required Zn++or Mn++for activity, was inhibited by heparin, and had its pH optimum at 6.0. Fraction E required Zn++for activity, was not inhibited by heparin, and had its pH optimum at 5.5. Fractions B, C, and D did not require metal ions for activity, and were distinguished from each other by differences in response to pH, cysteine, ethylenediaminetetraacetate, fluoride, and tartrate.The pH range over which fraction A was active was shifted to slightly higher values when Mn++was the activator rather than Zn++. Also, A was inhibited strongly by cysteine when activated by Zn++, but not when activated by Mn++. Data are presented that indicate these differences were due to different properties of the activating ions, rather than to the presence in fraction A of two enzymes, one activated by Zn++and the other by Mn++.

LINGCOD MUSCLE NUCLEASE

1959 ◽  
Vol 37 (8) ◽  
pp. 945-952 ◽  
Author(s):  
Neil Tomlinson
Keyword(s):  
Ribonucleic Acid ◽  
Free Base ◽  
Ophiodon Elongatus ◽  
Base Form ◽  
Nitrophenyl Phosphate ◽  
Diethylaminoethyl Cellulose ◽  
Cyclic Phosphates ◽  
Low Rate

A nuclease from muscle of lingcod (Ophiodon elongatus) has been purified by chromatography on diethylaminoethyl cellulose in the free-base form by stepwise elution with increasing concentrations of tris(hydroxymethyl)aminomethane. HCl, pH 7.0. The nuclease has been shown to hydrolyze ribonucleic acid, adenosine 3′-benzyl phosphate, thymidine 3′-p-nitrophenyl phosphate, and thymidine 5′-p-nitrophenyl phosphate. The latter compound was hydrolyzed at a very low rate. It did not hydrolyze adenosine 2′- or 5′- benzyl phosphate or certain nucleoside cyclic phosphates. The enzyme was inhibited by monoiodoacetate but not by heparin.

LINGCOD MUSCLE NUCLEASE

1959 ◽  
Vol 37 (1) ◽  
pp. 945-952 ◽  
Author(s):  
Neil Tomlinson
Keyword(s):  
Ribonucleic Acid ◽  
Free Base ◽  
Ophiodon Elongatus ◽  
Base Form ◽  
Nitrophenyl Phosphate ◽  
Diethylaminoethyl Cellulose ◽  
Cyclic Phosphates ◽  
Low Rate

A nuclease from muscle of lingcod (Ophiodon elongatus) has been purified by chromatography on diethylaminoethyl cellulose in the free-base form by stepwise elution with increasing concentrations of tris(hydroxymethyl)aminomethane. HCl, pH 7.0. The nuclease has been shown to hydrolyze ribonucleic acid, adenosine 3′-benzyl phosphate, thymidine 3′-p-nitrophenyl phosphate, and thymidine 5′-p-nitrophenyl phosphate. The latter compound was hydrolyzed at a very low rate. It did not hydrolyze adenosine 2′- or 5′- benzyl phosphate or certain nucleoside cyclic phosphates. The enzyme was inhibited by monoiodoacetate but not by heparin.

scholarly journals MICROSOMAL AND LYSOSOMAL ACID PHOSPHATASE ISOENZYMES OF MOUSE KIDNEY CHARACTERIZATION AND SEPARATION

1972 ◽  
Vol 20 (7) ◽  
pp. 487-498 ◽  
Author(s):  
CHI-WEI LIN ◽  
WILLIAM H. FISHMAN
Keyword(s):  
Acid Phosphatase ◽  
Migration Rate ◽  
Biochemical Properties ◽  
Mouse Kidney ◽  
Acid Phosphatases ◽  
Ph Optimum ◽  
Lysosomal Acid ◽  
Lysosomal Fraction ◽  
Heat Stable ◽  
Diethylaminoethyl Cellulose

Biochemical methods have been employed to characterize and separate acid phosphatases from lysosomal and microsomal fractions in order to decide whether different isoenzymes reside in these subcellular locations. Microsomal and lysosomal fractions of mouse kidney homogenate were isolated by differential centrifugation. Acid phosphatase of lysosomal fraction goes into solution after lysosomes have been repeatedly frozen and thawed, whereas acid phosphatase of microsomal fraction is firmly bound to the membrane and is freed of contamination by lysosomal enzyme after ultrasonication and centrifugation. The membrane-bound microsomal acid phosphatase is labile at 37°C, pH 4.9, more active toward phenolic substrates (phenyl phosphate and p-nitrophenyl phosphate) than β-glycerophosphate, α-naphthol phosphate or naphthol AS-BI phosphate. It also has a higher pH optimum (6.3), is resistant to l-tartrate and oxalate inhibition and has a slower electrophoretic migration rate in Triton X-100-impregnated polyacrylamide gels. The free lysosomal acid phosphatase is relatively heat-stable, is less active against phenolic substrates, is sensitive to l-tartrate and oxalate inhibition, has a lower pH optimum (5.6) and has a faster migration rate in electrophoresis. These two acid phosphatases can also be separated by diethylaminoethyl-cellulose chromatography. This study thus demonstrated the existence of an acid phosphatase isoenzyme in the microsomal membrane with different biochemical properties from the lysosomal isoenzyme of acid phosphatase.

scholarly journals The 110-kD protein-calmodulin complex of the intestinal microvillus is an actin-activated MgATPase.

1987 ◽  
Vol 105 (1) ◽  
pp. 313-324 ◽  
Author(s):  
K A Conzelman ◽  
M S Mooseker
Keyword(s):  
Enzymatic Activity ◽  
Nucleoside Triphosphate ◽  
Dependent Manner ◽  
Binding Interaction ◽  
Ph Optimum ◽  
Concentration Dependent Manner ◽  
Nitrophenyl Phosphate ◽  
Ph Range ◽  
Mgatpase Activity ◽  
Actin Concentration

The microvillus 110-kD protein-calmodulin complex (designated 110K-CM) shares several properties with all myosins. In addition to its well-defined ATP-dependent binding interaction with F-actin, 110K-CM is an ATPase with diagnostically myosin-like divalent cation sensitivity. It exhibits maximum enzymatic activity in the presence of K+ and EDTA (0.24 mumol P1/mg per min) or in the presence of Ca++ (0.40 mumol P1/mg per min) and significantly less activity in physiological ionic conditions of salt and Mg++ (0.04 mumol P1/mg per min). This MgATPase is activated by F-actin in an actin concentration-dependent manner (up to 2.5-3.5-fold). The specific MgATPase activity of 110K-CM is also enhanced by the addition of 5-10 microM Ca++, but in the isolated complex, there is often also a decrease in the extent of actin activation in this range of free Ca++. Actin activation is maintained, however, in samples with exogenously added calmodulin; under these conditions, there is an approximately sevenfold stimulation of 110K-CM's enzymatic activity in the presence of 5-10 microM Ca++ and actin. 110K-CM is relatively indiscriminant in its nucleoside triphosphate specificity; in addition to ATP, GTP, CTP, UTP, and ITP are all hydrolyzed by the complex in the presence of either Mg++ or Ca++. Neither AMP nor the phosphatase substrate p-nitrophenyl phosphate are substrates for the enzymatic activity. The pH optimum for CaATPase activity is 6.0-7.5; maximum actin activation of MgATPase occurs over a broad pH range of 6.5-8.5. Finally, like myosins, purified 110K-CM crosslinks actin filaments into loosely ordered aggregates in the absence of ATP. Collectively these data support the proposal of Collins and Borysenko (1984, J. Biol. Chem., 259:14128-14135) that the 110K-CM complex is functionally analogous to the mechanoenzyme myosin.

scholarly journals Heterogeneity of liver acid phosphatases in developing chick embryo

1969 ◽  
Vol 115 (2) ◽  
pp. 191-197 ◽  
Author(s):  
K.-M. Wang
Keyword(s):  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Soluble Fraction ◽  
The Other ◽  
Acid Phosphatases ◽  
Peak Activity ◽  
Ph Optimum ◽  
Triton X ◽  
Soluble Fractions

1. The development, localization and heterogeneity of acid phosphatase and a Zn2+-activated acid phosphatase in cellular fractions of developing chick liver were studied. 2. Acid phosphatase is distributed abundantly in the particulate and soluble fractions. The soluble fraction is rich in Zn2+-activated acid phosphatase, which attains its peak activity at about 15 days of incubation. 3. The particulate acid phosphatase activity is inhibited by fluoride but not by sodium l(+)-tartrate or cysteine. On the other hand, the soluble Zn2+-activated acid phosphatase activity is inhibited by sodium l(+)-tartrate and cysteine but not by fluoride. 4. The pH optimum of these two enzymes is similar at about 5·6. 5. The soluble Zn2+-activated acid phosphatase activity appears to be thermally stabilized by the treatment with Triton X-100 or bovine serum albumin.

The acid phosphatases of bovine leucocytes, plasma and the milk of healthy and mastitic cows

1975 ◽  
Vol 42 (3) ◽  
pp. 391-400 ◽  
Author(s):  
A. T. Andrews ◽  
E. Alichanidis
Keyword(s):  
Molecular Weight ◽  
Acid Phosphatase ◽  
Exchange Resin ◽  
Bovine Milk ◽  
Ion Exchange Resin ◽  
Acid Phosphatases ◽  
Ph Optimum ◽  
Nitrophenyl Phosphate ◽  
Competitive Inhibitors ◽  
Electrophoretic Component

SummarySome of the acid phosphatase isozymes of bovine leucocytes and plasma have been separated and partly characterized. About 80% of the phosphatase activity of leucocytes at pH 4·9 was particle-bound and about 8% was extractable with Amberlite CG-50 ion exchange resin. This extractable enzyme existed as a single electrophoretic component with a mol. wt of about 42000 and with optimum activity at pH 5·8. Km for p-nitrophenyl phosphate was 1·6 mM at pH 5·8 and 0·4 mM at pH 4·9. At pH 5·8 orthophosphate (K1 = 1·5 mM) and pyrophosphate (Ki = 4·1 mM) were competitive inhibitors. The enzyme was also strongly inhibited by F−, Al3+, IO4− and S2032−. The enzyme which was not extractable with Amberlite was very heterogeneous with respect to molecular weight. At the pH optimum (4·9), Km for p-nitrophenyl phosphate was 0·4 mM and orthophosphate (K1 = 2·3 mM) and pyrophosphate (K1 = 2·1 mM) were competitive inhibitors. Other inhibitors included F−, Al3+, Hg2+, IO4− and tartrate. The enzyme extracted from plasma by Amberlite CG-50 treatment had properties similar to that extracted from leucocytes. Normal bovine milk contained a single acid phosphatase, but milk from cows with mastitis showed 3 electrophoretic isozyme bands, one being the same as in normal milk; the 2 additional bands were of leucocyte origin.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

1974 ◽  
Vol 31 (01) ◽  
pp. 072-085 ◽  
Author(s):  
M Kopitar ◽  
M Stegnar ◽  
B Accetto ◽  
D Lebez
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Isolation And Characterization ◽  
Ph Range ◽  
Inhibition Studies ◽  
Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

Removal of Trace Metals From Wastewater by Activated Carbon

1973 ◽  
Vol 8 (1) ◽  
pp. 110-121
Author(s):  
A. Netzer ◽  
J.D. Norman
Keyword(s):  
Activated Carbon ◽  
Trace Metals ◽  
Organic Compounds ◽  
The Other ◽  
Published Data ◽  
Ph Adjustment ◽  
Nickel Silver ◽  
Ph Range ◽  
Carbon Treatment ◽  
Cobalt Copper

Abstract The merits of activated carbon for removal of organic compounds from wastewater have been well documented in the literature. On the other hand there is a lack of published data on the use of activated carbon for the removal of trace metals from wastewater. Experiments were designed to assess the possibility that activated carbon treatment would remove aluminum, cadmium, chromium, cobalt, copper, iron, lead, manganese, mercury, nickel, silver and zinc from wastewater. All metals studied were tested over the pH range 3-11. Greater than 99.5% removal was achieved by pH adjustment and activated carbon treatment for most of the metals tested.

Efficient Preconcentration of Cu2+, Zn2+ and Fe3+ with an Agarose-Schiff Base Sorbent

2007 ◽  
Vol 72 (7) ◽  
pp. 908-916 ◽  
Author(s):  
Payman Hashemi ◽  
Hatam Hassanvand ◽  
Hossain Naeimi
Keyword(s):  
Schiff Base ◽  
Metal Ions ◽  
Metal Ion ◽  
Good Accuracy ◽  
Kinetic Studies ◽  
Sample Volume ◽  
Effects Of Ph ◽  
Glass Column ◽  
High Concentrations ◽  
Ph Range

Sorption and preconcentration of Cu2+, Zn2+ and Fe3+ on a salen-type Schiff base, 2,2'- [ethane-1,2-diylbis(nitrilomethylidyne)]bis(2-methylphenol), chemically immobilized on a highly crosslinked agarose support, were studied. Kinetic studies showed higher sorption rates of Cu2+ and Fe3+ in comparison with Zn2+. Half-times (t1/2) of 31, 106 and 58 s were obtained for sorption of Cu2+, Zn2+ and Fe3+ by the sorbent, respectively. Effects of pH, eluent concentration and volume, ionic strength, buffer concentration, sample volume and interferences on the recovery of the metal ions were investigated. A 5-ml portion of 0.4 M HCl solution was sufficient for quantitative elution of the metal ions from 0.5 ml of the sorbent packed in a 6.5 mm i.d. glass column. Quantitative recoveries were obtained in a pH range 5.5-6.5 for all the analytes. The volumes to be concentrated exceeding 500 ml, ionic strengths as high as 0.5 mol l-1, and acetate buffer concentrations up to 0.3 mol l-1 for Zn2+ and 0.4 mol l-1 for Cu2+ and Fe3+ did not have any significant effect on the recoveries. The system tolerated relatively high concentrations of diverse ions. Preconcentration factors up to 100 and detection limits of 0.31, 0.16 and 1.73 μg l-1 were obtained for Cu2+, Zn2+ and Fe3+, respectively, for their determination by a flame AAS instrument. The method was successfully applied to the metal ion determinations in several river water samples with good accuracy.

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