scholarly journals Organic pyrophosphates as substrates for human alkaline phosphatases

1967 ◽  
Vol 105 (3) ◽  
pp. 1307-1312 ◽  
Author(s):  
R. Helen Eaton ◽  
D W Moss
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Inorganic Phosphate ◽  
Starch Gel Electrophoresis ◽  
Alkaline Phosphatases ◽  
Starch Gel ◽  
Pyrophosphatase Activity ◽  
Small Intestinal ◽  
Hydrolysis Of ◽  
Single Enzyme

1. Purified human liver and small-intestinal alkaline orthophosphatases release inorganic phosphate at appreciable rates from a variety of organic pyrophosphate substrates. 2. The pyrophosphatase action is inhibited by Mg2+ ions at concentrations that activate the hydrolysis of orthophosphate substrates by these enzymes. 3. The results of mixed-substrate experiments, denaturation studies with heat or urea and starch-gel electrophoresis suggest that both orthophosphatase and pyrophosphatase activities are, in each preparation, properties of a single enzyme. 4. Intestinal phosphatase shows greater pyrophosphatase activity relative to orthophosphatase than the liver enzyme.

scholarly journals ENZYMES IN HOG KIDNEY HYDROLYZING AMINO ACID NAPHTHYLAMIDES

1966 ◽  
Vol 14 (4) ◽  
pp. 314-325 ◽  
Author(s):  
T. VANHA-PERTTULA ◽  
V. K. HOPSU ◽  
G. G. GLENNER
Keyword(s):  
Amino Acid ◽  
Substrate Specificity ◽  
Gel Electrophoresis ◽  
Metal Ion ◽  
Total Activity ◽  
Starch Gel Electrophoresis ◽  
Enzyme Preparations ◽  
Starch Gel ◽  
Hydrolysis Of ◽  
Hog Kidney

Hydrolysis of β-naphthylamides of a number of amino acids and dipeptides and of a number of di- and tripeptides by hog kidney homogenate and by fractions obtained by various fractionation procedures has been studied. The substrates were found to be split by a soluble, apparently sulfhydryl-dependent enzyme, and by a particle-bound, metal-activated enzyme. The former constituted only a small part of the total activity. The latter was subfractionated by starch gel electrophoresis into two fractions with identical characteristics. The soluble and particle-bound enzymes differed also in their substrate specificity. The latter enzyme was solubilized, partially purified, characterized by some modifier compounds and compared with enzyme preparations obtained by various fractionation procedures presented by other investigators. Thus enzyme showed ion-determined substrate specificity, i.e., hydrolysis of some of the amino acid naphthylamides was found to be activated by Co++ while the hydrolysis of others was inhibited by the same metal ion.

A QUANTITATIVE METHOD FOR THE DETERMINATION OF CHOLINESTERASE ACTIVITY AFTER STARCH-GEL ELECTROPHORESIS

1966 ◽  
Vol 44 (6) ◽  
pp. 953-955 ◽  
Author(s):  
H. S. Funnell ◽  
W. T. Oliver
Keyword(s):  
Gel Electrophoresis ◽  
Quantitative Method ◽  
Cholinesterase Activity ◽  
Competitive Inhibition ◽  
Starch Gel Electrophoresis ◽  
Final Solution ◽  
Natural Activity ◽  
Starch Gel ◽  
Hydrolysis Of

The method described in this paper is based upon hydrolysis of the starch gel by sodium hydroxide, which releases the dye. The dye is then taken up in an immiscible solvent, and the color determined spectrophotometrically. The method gave good reproducibility in studies both of natural activity and competitive inhibition without requiring any changes in the usual methods. The amount of dye in the final solution was related to the activity towards the dye-producing substrate of the enzyme present in the gel section.

scholarly journals N-acetyl-β-d-glucosaminidase in marmoset kidney, serum and urine

1978 ◽  
Vol 175 (3) ◽  
pp. 859-867 ◽  
Author(s):  
R J Pierce ◽  
R G Price ◽  
J S L Fowler
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Deae Cellulose ◽  
Heat Stability ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
Serum And Urine

N-Acetyl-beta-D-glucosaminidase activities were determined in homogenates of marmoset kidney, in serum and in urine by using the 4-methylumbelliferyl substrate. The enzyme activity was separated into several components by DEAE-cellulose ion-exchange chromatography, starch-gel electrophoresis and isoelectric focusing. The kidney contained two major forms of the enzyme, A and B, which had similar pH optima and Km values. The A-form bound to DEAE-cellulose at pH 6.8, migrated towards the anode on starch-gel electrophoresis and had a pI of 5.0. The B-form did not bind to DEAE-cellulose at pH 6.8, remained near the origin on starch-gel electrophoresis and had a pI of 7.64. The isoenzymes also differed in heat stability, the B-form being the more stable. Serum contained B-form activity and, in addition, two intermediate forms (I1 and I2) were loosely bound to DEAE-cellulose. The serum A-form activity was less firmly bound to DEAE-cellulose than was the tissue A-form and was designated As. Serum from a pregnant marmoset contained a form which may be analogous to the human P-isoenzyme. Urine contained only a small amount of B-form activity, the majority being present in the A-form. The kidney A- and B-forms both had mol.wts. of 96000–100000 and the activity was predominantly lysosomal. Partial purification of the kidney A isoenzyme was undertaken. Immunoprecipitation studies indicated a relationship between marmoset kidney A-form and human liver A-form activity.

scholarly journals MOUSE ERYTHROCYTE ESTERASES AND AN INTERACTION WITH HEPARIN AS SHOWN BY STARCH GEL ELECTROPHORESIS

1969 ◽  
Vol 17 (2) ◽  
pp. 95-101 ◽  
Author(s):  
McCORMICK TEMPLETON
Keyword(s):  
Young Adult ◽  
Enzymatic Hydrolysis ◽  
Adult Male ◽  
Gel Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Acetate Esters ◽  
Mouse Erythrocyte ◽  
Starch Gel ◽  
Hydrolysis Of ◽  
Naphthyl Acetate

Esterases from the blood of young adult male C-57 brown mice were separated by starch gel electrophoresis. Enzymatic hydrolysis of the substrates α-naphthyl acetate, naphthyl-AS acetate, α-naphthyl butyrate and acetylthiocholine was studied. Three different esterases were found in erythrocytes. One band hydrolyzed a-naphthyl butyrate but was only slightly active with acetate esters. The fastest migrating bands hydrolyzed acetate faster than butyrate esters, but would hydrolyze both substrates. A third esterase represented in the zymogram by a pair of bands (called "acetate" bands) would hydrolyze only acetate esters. When heparin, or plasma containing heparin, was inserted in a gel so as to overlap an erythrocyte sample, only the acetate bands were altered. When hepanin was inserted cathodal to an erythrocyte sample, the acetate bands are selectively affected by the heparin in such a way as to cause the acetate band to migrate more rapidly toward the anode; other esterase bands were not similarly affected. Acetate bands were not inhibited by physostygmine (1 mM) but their activities diminished with increased concentrations of buffer.

Fractionation of Plasminogen by Starch Gel Electrophoresis

1964 ◽  
Vol 12 (01) ◽  
pp. 126-136 ◽  
Author(s):  
Karl H. Slotta ◽  
J. D Gonzalez
Keyword(s):  
Gel Electrophoresis ◽  
Room Temperature ◽  
Broad Band ◽  
Caproic Acid ◽  
Starch Gel Electrophoresis ◽  
Full Activity ◽  
Veronal Buffer ◽  
Starch Gel ◽  
Alkaline Range

SummaryWhen urea or ε-amino caproic acid were used as solublizing agents for plasminogen in electrophoretic experiments, only one broad band of the proenzyme was obtained on acetate cellulose, in starch block, and in acrylamide gel. In starch gel electrophoresis, however, both forms of plasminogen – the native or euglobulin and Kline’s or Pseudoglobulin plasminogen – separated into six bands. These migrated toward the cathode at room temperature in borate or veronal buffer in the alkaline range and showed full activity in fibrinagar-streptokinase plates.

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