scholarly journals Studies on the properties of cow's-milk tributyrinases and their interaction with milk proteins

1966 ◽  
Vol 101 (3) ◽  
pp. 651-660 ◽  
Author(s):  
WK Downey ◽  
P Andrews
Keyword(s):  
Molecular Weight ◽  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Sodium Chloride ◽  
Xanthine Oxidase ◽  
Pancreatic Lipase ◽  
Wheat Germ ◽  
Milk Proteins ◽  
Casein Micelles ◽  
Bivalent Cation

1. The tributyrinases in milk are mainly associated with casein micelles. Dilution or addition of sodium chloride increases the enzyme activity, probably by dissociating the micelle-tributyrinase complexes. 2. Tributyrinase activities of milks activated by dilution and sodium chloride addition were in the range 0.2-1.7muequiv. of acid liberated/ml. of milk/min. from tributyrin emulsion at pH8.5 and 25 degrees . The enzymes have a bivalent-cation requirement for full activity and are rather unstable when separated from casein. 3. Ultracentrifugation of skim milks containing sodium chloride (0.75m) gave preparations low in casein but containing about 70% of the milk tributyrinases. The tributyrinases in such preparations appear to be bound in complexes of molecular weight about 350000. Dilution may result in dissociation to give the free enzymes. 4. Pancreatic lipase also formed complexes with casein micelles, but wheat-germ esterase, xanthine oxidase, milk alkaline phosphatase and other enzymes did not.

Association of lipases with micellar and soluble casein complexes

1970 ◽  
Vol 37 (1) ◽  
pp. 47-59 ◽  
Author(s):  
W. K. Downey ◽  
R. F. Murphy
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Calcium Phosphate ◽  
Pancreatic Lipase ◽  
Gel Filtration ◽  
Casein Micelles ◽  
Micellar Structure ◽  
Milk Serum ◽  
Colloidal Calcium Phosphate ◽  
Soluble Complexes

SummaryThe association of lipase with casein micelles and soluble casein complexes was investigated by gel-filtration on Sephadex G-200 and Sepharose 2B columns which were equilibrated with synthetic milk serum. Gel-filtration indicated that the molecular weight of casein micelles in milk is > 108 whereas the casein in colloidal phosphate-free milk is present as soluble complexes of molecular weight ca. 2×106 containing αs-, β- and κ-casein. The soluble complexes appear to be stabilized in the micelle by colloidal calcium phosphate linkages. On addition of pancreatic lipase to milk, activity was impaired due to binding of the enzyme both to micellar and to soluble casein complexes. The enzyme dissociated from the latter during gel-filtration on Sepharose 2B columns. The binding of lipase to casein was not dependent on the presence of colloidal phosphate and consequently complete micellar structure is not essential for association of lipase with casein. Binding of the lipase to phosvitin did not result in a loss of enzyme activity. Lipases in milk appear to be involved in the equilibrium between micellar and soluble casein. The activity of lipases in milk is apparently influenced by this equilibrium. Some problems encountered in the use of gel-filtration to study the interactions of lipases with caseins are described.

Extracellular alkaline phosphatase from marine bacteria: purification and properties of extracellular phosphatase from a marine Pseudomonas sp.

1980 ◽  
Vol 26 (7) ◽  
pp. 833-838 ◽  
Author(s):  
Hiromi Kobori ◽  
Nobuo Taga
Keyword(s):  
Molecular Weight ◽  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Marine Bacteria ◽  
Divalent Cations ◽  
Maximal Activity ◽  
Pseudomonas Sp ◽  
Purification And Properties ◽  
Extracellular Alkaline Phosphatase ◽  
Increased Pressure

Extracellular alkaline phosphatase produced by a marine Pseudomonas was purified to electrophoretic homogeneity. The molecular weight of the enzyme was estimated to be 100 000. The enzyme had maximal activity at pH 11.5. The enzyme was completely inhibited by 1 mM EDTA. However, divalent cations reversed the enzyme inhibition and their order of effectiveness on the reaction was Zn2+ > Ca2+ > Mn2+ > Mg2+ > Sr2+ > Co2+. The enzyme activity was affected by the species of anion whose order of effectiveness was demonstrated to follow the lyotrophic series, Cl− > Br− > NO3−> ClO4− > SCN−. The activity of phosphatase was accelerated linearly by increased pressure until up to 1000 atm (1 atm = 101.325 kPa), and the enzyme activity at 1000 atm was 3.2 times higher than that at 1 atm.

FREEZING DAMAGE TO BOVINE CREAM INDICATED BY RELEASE OF ENZYMES

1959 ◽  
Vol 37 (7) ◽  
pp. 821-827 ◽  
Author(s):  
L. J. N. Cole ◽  
D. Kluepfel ◽  
C. V. Lusena
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Xanthine Oxidase ◽  
Aqueous Phase ◽  
The Other ◽  
Rapid Freezing ◽  
Freezing Damage ◽  
Membrane Material ◽  
Fat Globules ◽  
Main Effect

When washed cream was frozen slowly and thawed, some breaking of the emulsion occurred and on centrifugation a pellet, mostly membrane material, was obtained. Xanthine oxidase and alkaline phosphatase were present in this pellet, but little enzyme activity was found in the aqueous phase. The main effect of slow freezing was to force the fat globules together so that alteration and redistribution of the membranes could occur, and, on thawing, fat could coalesce. Rapid freezing on the other hand distributed fat globules more evenly so that less coalescence could occur on thawing.

FREEZING DAMAGE TO BOVINE CREAM INDICATED BY RELEASE OF ENZYMES

1959 ◽  
Vol 37 (1) ◽  
pp. 821-827 ◽  
Author(s):  
L. J. N. Cole ◽  
D. Kluepfel ◽  
C. V. Lusena
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Xanthine Oxidase ◽  
Aqueous Phase ◽  
The Other ◽  
Rapid Freezing ◽  
Freezing Damage ◽  
Membrane Material ◽  
Fat Globules ◽  
Main Effect

When washed cream was frozen slowly and thawed, some breaking of the emulsion occurred and on centrifugation a pellet, mostly membrane material, was obtained. Xanthine oxidase and alkaline phosphatase were present in this pellet, but little enzyme activity was found in the aqueous phase. The main effect of slow freezing was to force the fat globules together so that alteration and redistribution of the membranes could occur, and, on thawing, fat could coalesce. Rapid freezing on the other hand distributed fat globules more evenly so that less coalescence could occur on thawing.

Effects of Metal Poisoning on Five Liver Enzymes in the Killifish (Fundulus heteroclitus)

1970 ◽  
Vol 27 (2) ◽  
pp. 383-390 ◽  
Author(s):  
Eugene Jackim ◽  
Janice M. Hamlin ◽  
Stephen Sonis
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Xanthine Oxidase ◽  
Liver Enzyme ◽  
Fundulus Heteroclitus ◽  
Liver Enzymes ◽  
Enzyme Preparations ◽  
Liver Enzyme Activity ◽  
Acid And Alkaline Phosphatase ◽  
Metal Poisoning

Activities of five liver enzymes (acid and alkaline phosphatase, catalase, xanthine oxidase, and ribonuclease) from Fundulus heteroclitus surviving exposure to 96-hr TLm concentrations of salts of six metals (lead, copper, mercury, beryllium, cadmium, and silver) differed markedly from those of unexposed fish. Changes in enzyme activity produced by the exposures were not necessarily the same in magnitude or direction as those observed when the salts were introduced directly into the enzyme preparations. It is proposed that changes in liver enzyme activity may be useful as a kind of biochemical autopsy tool for diagnosing sublethal metal poisoning in fish.

scholarly journals Guinea-pig milk-protein synthesis. Isolation and characterization of messenger ribonucleic acids from lactating mammary gland and identification of caseins and pre-α-lactalbumin as translation products in heterologous cell-free systems

1976 ◽  
Vol 160 (1) ◽  
pp. 57-74 ◽  
Author(s):  
R K Craig ◽  
P A Brown ◽  
O S Harrison ◽  
D McIlreavy ◽  
P N Campbell
Keyword(s):  
Molecular Weight ◽  
Protein Synthesis ◽  
Mammary Gland ◽  
Guinea Pig ◽  
Wheat Germ ◽  
Milk Proteins ◽  
N Terminus ◽  
Lactating Mammary Gland ◽  
Heterologous Cell ◽  
Alpha Lactalbumin

1. The major milk proteins synthesized by the lactating mammary gland of the guinea pig were identified and designated as caseins A, B and C and alpha-lactalbumin, with estimated mol.wts. of 28000, 25500, 20500 and 14500 respectively. 2. Antisera to the total casein fraction and to alpha-lactalbumin were prepared from rabbits. The milk proteins were also iodinated with either 131I or 125I. 3. A poly(A)-rich RNA fraction was isolated from lactating guinea-pig mammary glands. Isolation was by affinity chromatography on oligo(dT)-cellulose. 4. Examination of this RNA fraction by electrophoresis on polyacrylamide gels containing formamide indicated three major species 1, 2 and 3, with estimated wol.wts. of 5.4 × 10(5) and 3.3 × 10(5), and the apparent absence of rRNA species. 5. The poly(A)-rich RNA stimulated protein synthesis in heterologous cell-free systems based on wheat germ, Krebs II ascites-tumour cells, and the latter supplemented with an initiation factor-3 fraction from rabbit reticulocyte ribosomes. 6. Between 80 and 90% of the protein synthesis directed by the mRNA was for milk proteins. 7. Analysis of the proteins immunoprecipitated by the alpha-lactalbumin antiserum showed in the wheat-germ system that the product was a protein with a molecular weight greater than that of alpha-lactalbumin, whereas in the ascites-tumour-cell systems both this protein and alpha-lactalbumin were found. When the larger protein was treated with CNBr and the resulting peptides were examined, it was shown that the extra peptide was at the N-terminus. This and other evidence is adduced for the initial translation product of alpha-lactalbumin being a precursor with an addition of about ten amino acids at the N-terminus. 8. Similar analysis of the casein immlnospecific proteins produced under the direction of mRNA indicated that the products had a molecular weight that was apparently a littel smaller than that of the caseins synthesized in vivo. This was not consistent with higher-molecular weight casein precursors. 9. Possible explanations for the results obtained are discussed, especially in terms of the physiological significance of the pre-alpha-lactalbumin as a secretory protein.

Formation and Structure of Casein Micelles in Lactating Mammary Tissue

1970 ◽  
Vol 28 ◽  
pp. 150-151
Author(s):  
Robert J. Carroll ◽  
Marvin P. Thompson ◽  
Harold M. Farrell
Keyword(s):  
Size Distribution ◽  
G Protein ◽  
Milk Proteins ◽  
Mammary Tissue ◽  
Colloidal System ◽  
Casein Micelles ◽  
The Stability

Milk is an unusually stable colloidal system; the stability of this system is due primarily to the formation of micelles by the major milk proteins, the caseins. Numerous models for the structure of casein micelles have been proposed; these models have been formulated on the basis of in vitro studies. Synthetic casein micelles (i.e., those formed by mixing the purified αsl- and k-caseins with Ca2+ in appropriate ratios) are dissimilar to those from freshly-drawn milks in (i) size distribution, (ii) ratio of Ca/P, and (iii) solvation (g. water/g. protein). Evidently, in vivo organization of the caseins into the micellar form occurs in-a manner which is not identical to the in vitro mode of formation.

Exterior and Interior Events in Early Stage of Thrombin-induced Platelet Aggregation

1977 ◽  
Vol 38 (03) ◽  
pp. 0630-0639 ◽  
Author(s):  
Shuichi Hashimoto ◽  
Sachiko Shibata ◽  
Bonro Kobayashi
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Plasma Membrane ◽  
Platelet Aggregation ◽  
Early Stage ◽  
Adenyl Cyclase ◽  
Cellular Component ◽  
Primary Event ◽  
Rabbit Platelets ◽  
Membrane Bound

SummaryTreatment of washed rabbit platelets with 1 u/ml of thrombin at 37° C resulted in a disappearance from platelets of a protein with 250,000 dalton molecular weight which was shown to be originated from plasma membrane. Parallel loss of adenyl cyclase was noted, and both reactions were complete within 30 sec. From the patterns of disc electrophoretograms, the importance of quick suppression of thrombin action in demonstrating the primary event was stressed.Thrombin induced an apparent activation of membrane bound phosphodiesterase. This reaction was also complete within 30 sec. The cellular component which contained the enzyme activity was distinct from plasma membrane. Soluble phosphodiesterase was not influenced by thrombin at all.These reactions required intact platelet cells to react with thrombin, and no reaction was detected when subcellular preparations were treated with thrombin.Possibility of collaboration of changes in externally located synthetic enzyme with those in internally located degrading enzyme in the early phase of thrombin action on platelets was suggested.

scholarly journals Inhibition studies of alkaline phosphatase in hard tissue-forming cells.

1975 ◽  
Vol 23 (5) ◽  
pp. 342-347 ◽  
Author(s):  
A Linde ◽  
B C Magnusson
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Adenosine Triphosphatase ◽  
Ruthenium Red ◽  
Inorganic Pyrophosphatase ◽  
Alkaline Ph ◽  
Hard Tissue ◽  
Assay Condition ◽  
Phosphatase Inhibitors ◽  
Inhibition Studies

The effects of the alkaline phosphatase inhibitors levamisole and R 8231 on p-nitro-phenylphosphatase, inorganic pyrophosphatase and adenosine triphosphatase (ATPase) activities in dentingenically active odontoblasts were studied. The p-nitrophenylphosphatase and inorganic pyrophosphatase activities were inhibited, while 40% of the ATP-splitting enzyme activity remained under the assay condition used. This finding, togeather with earlier studies, indicates that at least two different phosphatase are active at alkaline pH in hard tissue-forming cells; on nonspecific alkaline phosphatase and one specific ATPase. The ATPase activity is uninfluenced by ouabain and ruthenium red and is activated by Ca-2+ ions.

Occurrence of milk proteins in the urine of cows during an extended milking interval

1967 ◽  
Vol 34 (1) ◽  
pp. 27-30 ◽  
Author(s):  
R. L. J. Lyster ◽  
J. V. Wheelock
Keyword(s):  
Molecular Weight ◽  
Milk Proteins ◽  
Immunological Methods ◽  
Factors Affecting ◽  
Β Lactoglobulin

SummaryImmunological methods have been used to test samples of urine from 5 cows for the presence of milk proteins. None could be detected when the cows were milked twice daily at the usual intervals, but during an extended milking interval α-lactalbumin was found in the urine of all 5 cows and β-lactoglobulin in the urine of 2 cows. The urine of one cow during and after a milking interval of 39 h contained 1·63 g α-lactalbumin, 1·12 g β-lactoglobulin and a small amount of casein. One of the factors affecting the transfer of these milk constituents from the udder to the urine appears to be their molecular weight.

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