HYDROLYTIC ENZYMES IN BOVINE SKELETAL MUSCLE: I. STARCH GEL ELEGTROPHORETIC SEPARATION AND PROPERTIES OF SOLUBLE ESTERASES

1965 ◽  
Vol 43 (11) ◽  
pp. 1779-1786 ◽  
Author(s):  
H. F. MacRae ◽  
C. J. Randall
Keyword(s):  
Skeletal Muscle ◽  
Skeletal Muscles ◽  
Hydrolytic Enzymes ◽  
Water Soluble ◽  
Heat Stable ◽  
Heat Labile ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Bovine Skeletal Muscle ◽  
Naphthyl Acetate

Water-soluble esterases of certain bovine skeletal muscles were separated by horizontal zone electrophoresis in starch gel in a discontinuous ouffer system. Eighteen bands of esterase activity were detected by the use of α-naphthyl acetate and α-naphthyl butyrate as substrates. Other substrates and various inhibitors were used to characterize the separated enzymes. A group of presumed isozymic esterases (three bands), which hydrolyzed α-naphthyl butyrate but not any other substrate tested, was sensitive to organophosphates, was heat labile, and was classified as aliesterase or, more specifically, as butyrylesterase. Another group of presumed isozymic esterases (four bands) hydrolyzed only α-naphthyl acetate and indoxyl acetates, was heat stable and resistant to organophosphates, and was tentatively classified as arylesterase or cathepsin. Eleven heat-labile esterase bands hydrolyzed both α-naphthyl acetate and α-naphthyl butyrate, were sensitive to organophosphates, and were classified as nonspecific aliesterases.

PROPERTIES AND CLASSIFICATION OF THE SOLUBLE ESTERASES OF HUMAN BRAIN

1966 ◽  
Vol 44 (2) ◽  
pp. 225-232 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Alkaline Phosphatase ◽  
Human Brain ◽  
Esterase Activity ◽  
Water Soluble ◽  
Human Tissues ◽  
Esterase Pattern ◽  
Vertical Zone ◽  
Zone Electrophoresis ◽  
Starch Gel

Water-soluble proteins and enzymes of human brain were separated by vertical zone electrophoresis in starch gel. Fifteen bands of esterase activity were detected in brain. Various substrates and inhibitors were used in efforts to identify enzymes in addition to a comparison of the esterase pattern with patterns obtained from other human tissues. One zone, composed of four bands of acetylesterase activity, was found to be common to all the tissues investigated with the exception of serum. Two bands of cholinesterase and two bands of A-esterase activity were identified. The remaining bands, which were aliesterases possessing broad overlapping substrate specificity and inhibitor sensitivity, were electrophoretically different from those of other tissues. Observations on alkaline phosphatase, acid phosphatase, and lactate dehydrogenase were recorded for comparison with the data on esterases.

PROPERTIES AND CLASSIFICATION OF THE SOLUBLE ESTERASES OF HUMAN SKELETAL AND SMOOTH MUSCLE

1965 ◽  
Vol 43 (1) ◽  
pp. 73-79 ◽  
Author(s):  
D. J. Ecobichon ◽  
W. Kalow
Keyword(s):  
Skeletal Muscle ◽  
Smooth Muscle ◽  
Human Liver ◽  
Renal Tissue ◽  
Water Soluble ◽  
Serum Cholinesterase ◽  
Liver And Kidney ◽  
Starch Gel ◽  
Zone Characteristic

Water-soluble proteins and enzymes of human skeletal and smooth muscle were separated by vertical-zone electrophoresis in starch gel and compared with those of human liver and kidney. Thirteen bands of proteins were detected with amido black in skeletal muscle, five of which were also detected in smooth muscle. Various substrates and inhibitors were used in efforts to identify enzymes. Ten bands of esterase activity were detected in skeletal muscle, and nine in smooth muscle. One zone, characteristic of serum cholinesterase, was believed to be due to serum contained in the tissue. A zone of isozymic esterases found in skeletal and smooth muscle was similar to a zone in human liver and kidney and reacted like an acetylesterase. Other esterase bands, which showed a marked hydrolysis of α-naphthyl butyrate, were similar to aliesterases of renal tissue. Observations on alkaline phosphatase, acid phosphatase, aminopeptidase, lactate dehydrogenase, and catalase were recorded for comparison with the data on esterases.

The water-soluble proteins of bovine skeletal muscle

1960 ◽  
Vol 86 (2) ◽  
pp. 238-250 ◽  
Author(s):  
M.J. Kronman ◽  
L.E. Weinberger ◽  
R.J. Winterbottom
Keyword(s):  
Skeletal Muscle ◽  
Soluble Proteins ◽  
Water Soluble ◽  
Bovine Skeletal Muscle

SOME PROPERTIES OF THE SOLUBLE ESTERASES OF HUMAN LIVER

1961 ◽  
Vol 39 (9) ◽  
pp. 1329-1332 ◽  
Author(s):  
D. J. Ecobichon ◽  
W. Kalow
Keyword(s):  
Human Serum ◽  
Human Liver ◽  
Histochemical Staining ◽  
Enzymatic Properties ◽  
Water Soluble ◽  
Electrophoretic Migration ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Staining Methods

Zone electrophoresis on starch gel in conjunction with various histochemical staining methods was applied to the study of the water-soluble esterases of liver. The results indicated that in regard to electrophoretic migration and enzymatic properties, none of the human liver esterases was identical with any of the human serum esterases.

CHARACTERIZATION OF THE ESTERASES FROM ELECTRIC TISSUE OF ELECTROPHORUS BY STARCH-GEL ELECTROPHORESIS

1967 ◽  
Vol 45 (7) ◽  
pp. 1099-1105 ◽  
Author(s):  
D. J. Ecobichon ◽  
Y. Israel
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Water Soluble ◽  
Starch Gel Electrophoresis ◽  
Electrophorus Electricus ◽  
Vertical Zone ◽  
Zone Electrophoresis ◽  
Starch Gel

The water-soluble esterases of a microsome-free supernatant of the electric tissue of Electrophorus electricus were separated by vertical-zone electrophoresis in starch gel. Specific and nonspecific substrates and inhibitors were used in conjunction with histochemical techniques to identify the enzymes. Acetylcholinesterase was present in the form of four bands of activity, the electrophoretic mobility of which was suggestive of aggregated forms of the enzyme. Pseudocholinesterase was detected as two weak bands of activity. A third esterase was identified as a nonspecific carboxylesterase and shown to be a sialoprotein.

MULTIPLE FORMS OF HUMAN LIVER ESTERASES

1965 ◽  
Vol 43 (5) ◽  
pp. 595-602 ◽  
Author(s):  
D. J. Ecobichon
Keyword(s):  
Human Liver ◽  
Molecular Size ◽  
Water Soluble ◽  
Electrical Charge ◽  
Multiple Forms ◽  
Substrate Specificities ◽  
Graphic Analysis ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Relative Retardation

Zone electrophoresis in starch gel of the water-soluble human liver esterases resulted in the separation of three zones of activity, each composed of several bands. The relative sizes of the enzymes in each zone were studied by utilizing the relative retardation of the electrophoretic migration induced by changes in the concentration of starch. On the basis of graphic analysis, four esterase bands comprising the zone migrating towards the cathode were found to be similar in molecular size or shape. A similar observation was made for seven bands comprising a zone migrating towards the anode. Taken with the substrate specificities and sensitivities toward various inhibitors, these observations strengthen the hypothesis that at least two of the hepatic esterases, an acetylesterase and an aliesterase, exist as multiple forms, differing primarily in net electrical charge.

scholarly journals Paroxysmal Nocturnal Hemoglobinuria

Blood ◽  
1953 ◽  
Vol 8 (5) ◽  
pp. 444-458 ◽  
Author(s):  
WILLIAM H. CROSBY
Keyword(s):  
Hemolytic Activity ◽  
Soluble Fraction ◽  
Insoluble Fraction ◽  
Water Soluble ◽  
Red Cells ◽  
Distilled Water ◽  
Heat Stable ◽  
Heat Labile

Abstract This report demonstrates the role and to some extent the interrelations of various factors that are active in the PNH hemolytic system. 1. Activity of four plasma factors, probably protein in nature, has been demonstrated. Two of these factors are hemolytic against PNH red cells, but not against normal red cells. The other two inhibit PNH hemolysis. (a). The heat labile hemolytic factor is water soluble and is therefore present in the soluble fraction of serum that has been dialyzed against distilled water. It is almost completely destroyed by heating at 53 C. for 10 minutes. It is slowly inactivated by incubation at 37 C. with 100 units per ml. of thrombin. It is rapidly destroyed by concentrations of thrombin in excess of 200 units per ml. It is inactive unless the heat stable hemolytic factor is also present. (b). The heat stable hemolytic factor is insoluble in water and is therefore precipitated from serum by dialysis against distilled water. It is quite resistant to 100 units per ml. of thrombin and to incubation at 53 C. It is inactive unless the heat labile hemolytic factor is also present. (c). The heat labile inhibitor is insoluble in water and is therefore found in the insoluble fraction of serum dialyzed against distilled water. It is inactivated by heating at 53 C. for 10 minutes but not by incubation with 100 units per ml. of thrombin. (d). The heat stable inhibitor is found in the water-soluble fraction of dialyzed serum. It withstands dialysis poorly, but it is not affected by 30 minutes incubation at 53 C. Incubation at 37 C. with 100 units per ml. of thrombin for 10 minutes destroys its inhibitory activity. Apparently the inhibitors are not interdependent. 2. Calcium in small amounts is probably essential to the PNH hemolytic system. The concentration of calcium that is optimum for hemolysis lies in the neighborhood of 2.5 mM. The optimum is a little less than the amount normally present in the plasma. Calcium in excess inhibits hemolysis in vitro, and no hemolysis occurs when the concentration exceeds 25 mM. per liter. 3. Magnesium is also essential to the PNH hemolytic system. As magnesium is added to the system in vitro hemolytic activity increases until the concentration exceeds 10 mM. per liter. Amounts greater than that have some dampening effect. Magnesium appears to antagonize the heat stable inhibitor of the PNH hemolytic system. 4. Thrombin is involved in this system insofar as the heat stable inhibitor and the heat labile hemolytic factor may be destroyed by thrombic activity. The inhibitor is rapidly destroyed, the hemolytic factor slowly. Therefore, the sum of the reaction to small amounts of thrombin in the PNH hemolytic system is to increase hemolytic activity. 5. Dilute heparin and protamine increase the activity of the PNH hemolytic system in vitro, probably by blocking the two inhibitors. Heparin appears to work against the heat stable inhibitor, protamine against the heat labile inhibitor. 6. The intensity of PNH hemolytic activity whether in vitro or in vivo is probably related to a balance that exists between the inhibitors and the hemolytic factors. Hemolytic crises may occur when the hemolytic factors are increased or when their antagonists are depressed.

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