CHARACTERISTICS OF AN EXTRACELLULAR PROTEINASE FROM MICROCOCCUS FREUDENREICHII

A proteolytic enzyme present in culture filtrates of Penicillium cyaneo-fulvum was purified approximately 100-fold. In the ultracentrifuge the enzyme behaved as a homogeneous protein, but on electrophoresis some contamination was apparent. The molecular weight of the enzyme was estimated to be about 45,000.The protease hydrolyzed casein and denatured haemoglobin, gelatin, and native bovine plasma albumin but not native or denatured ovalbumin. It also coagulated milk. The optimum pH for caseolysis was 9.5 to 11.0. Metal chelating- and sulphydryl-reagents did not affect enzyme activity but zinc and mercurous ions inhibited the enzyme, the inhibition being reversed with ethylenediaminetetraacetic acid (EDTA). Soybean trypsin inhibitor was without effect on the enzyme whereas ovomucoid inhibited the enzyme. Although it is similar in some respects to other alkaline proteases, the P. cyaneo-fulvum enzyme does not appear to be identical with any one of them.

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PURIFICATION AND PROPERTIES OF A PROTEASE FROM PENICILLIUM CYANEO-FULVUM

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o60-121 ◽

1960 ◽

Vol 38 (9) ◽

pp. 969-980 ◽

Cited By ~ 10

Author(s):

Kartar Singh ◽

S. M. Martin

Keyword(s):

Molecular Weight ◽

Enzyme Activity ◽

Proteolytic Enzyme ◽

Ethylenediaminetetraacetic Acid ◽

Alkaline Proteases ◽

Plasma Albumin ◽

Metal Chelating ◽

Bovine Plasma ◽

Purification And Properties ◽

Optimum Ph

A proteolytic enzyme present in culture filtrates of Penicillium cyaneo-fulvum was purified approximately 100-fold. In the ultracentrifuge the enzyme behaved as a homogeneous protein, but on electrophoresis some contamination was apparent. The molecular weight of the enzyme was estimated to be about 45,000.The protease hydrolyzed casein and denatured haemoglobin, gelatin, and native bovine plasma albumin but not native or denatured ovalbumin. It also coagulated milk. The optimum pH for caseolysis was 9.5 to 11.0. Metal chelating- and sulphydryl-reagents did not affect enzyme activity but zinc and mercurous ions inhibited the enzyme, the inhibition being reversed with ethylenediaminetetraacetic acid (EDTA). Soybean trypsin inhibitor was without effect on the enzyme whereas ovomucoid inhibited the enzyme. Although it is similar in some respects to other alkaline proteases, the P. cyaneo-fulvum enzyme does not appear to be identical with any one of them.

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THE EFFECT OF DIFFERENT PROTEIN SOLVENTS ON THE DOSE-RESPONSE LINE FOR THE INTRAVAGINAL ADMINISTRATION OF OESTRONE IN MICE

Journal of Endocrinology ◽

10.1677/joe.0.0090136 ◽

1953 ◽

Vol 9 (2) ◽

pp. 136-144 ◽

Cited By ~ 4

Author(s):

J. D. BIGGERS

Keyword(s):

Significant Influence ◽

Dose Response ◽

Significant Alteration ◽

Physical Interaction ◽

Plasma Albumin ◽

Egg Albumin ◽

Bovine Plasma ◽

Intravaginal Administration ◽

Response Line ◽

Β Lactoglobulin

1. The proteins: egg albumin and bovine plasma albumin exert a significant influence on the action of oestrone upon the vagina, but they differ in the concentrations required to produce an effect. Egg albumin does so only in 1·0 % solutions, while plasma albumin is effective in 0·01 % solutions. 2. β-Lactoglobulin may have an effect similar to that of egg albumin. 3. Globulins, fibrinogen, and pepsin have no influence on the action of oestrone upon the vagina. 4. While 1% egg albumin causes a significant alteration only in the position of the dose-response line, 0·01% plasma albumin, in addition, depresses the slope. 5. The significance of these results in relation to (a) the intravaginal dose-response line, and (b) the physical interaction of proteins and oestrogens is discussed.

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THE ROLE OF THE RETICULOENDOTHELIAL SYSTEM IN MOUSE ANAPHYLAXIS AS TESTED WITH HOMOLOGOUS ANTIBODY-ANTIGEN COMPLEXES

Journal of Experimental Medicine ◽

10.1084/jem.111.5.631 ◽

1960 ◽

Vol 111 (5) ◽

pp. 631-641 ◽

Cited By ~ 1

Author(s):

Richard Wistar ◽

Perry E. Treadwell ◽

A. F. Rasmussen

Keyword(s):

Intravenous Injection ◽

Reticuloendothelial System ◽

Soluble Antigen ◽

Plasma Albumin ◽

Carbon Particles ◽

Fresh Serum ◽

Bovine Plasma ◽

Antigen Antibody ◽

Mouse Antibody

Blockade of the RES was accomplished by the intravenous injection of carbon particles, thorotrast, zymosan, or a suspension of Bordetella pertussis. The blockade resulted in a decrease in sensitivity to anaphylaxis produced by the intravenous injection of soluble antigen-antibody complexes consisting of an optimal shocking mixture of bovine plasma albumin and mouse antibody to this antigen. The decrease in sensitivity to anaphylaxis was dependent on the dose of blockading agent and on the time between blockade and challenge with complex. The loss of sensitivity to anaphylaxis could not be restored by the administration of fresh serum from normal mice nor by guinea pig complement. Antigen-antibody complexes were rapidly removed from the blood with an average half-time of 11.9 minutes in normal mice. Complexes were cleared at significantly more rapid rates in mice previously sensitized to antigen. Although not all the results can be explained on the basis of blockade the facts indicate that the RES does play an important but as yet undefined role in passive homologous anaphylaxis in the mouse.

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Bovine Platelet Proteins

Thrombosis and Haemostasis ◽

10.1055/s-0038-1653555 ◽

1969 ◽

Vol 21 (03) ◽

pp. 409-418 ◽

Cited By ~ 2

Author(s):

S Łopaciuk ◽

N. O Solum

Keyword(s):

Bovine Serum ◽

Protein Composition ◽

Disc Electrophoresis ◽

Plasma Fibrinogen ◽

Plasma Albumin ◽

Precipitin Line ◽

Bovine Plasma ◽

Platelet Proteins

Summary1. The protein composition of bovine platelet extracts has been investigated by immunoelectrophoresis and polyacrylamide disc electrophoresis. The information obtained is discussed as a basis for study on platelet fibrinogen.2. With antiserum to platelet proteins 11 precipitin lines were observed 3 of which corresponded electrophoretically to plasma albumin, fibrinogen and γ-globulin. These lines were not seen using the same antiserum absorbed with bovine plasma. The 8 additional lines were still present indicating that they represented specific platelet components. Antiserum to plasma produced the 3 above-mentioned lines, but no others.3. With antiserum to purified bovine plasma fibrinogen 3 precipitin lines were observed. The fibrinogen line was the dominant one. The 2 additional lines did not disappear by absorption of the antiserum with bovine serum nor by incubation of the extracts with thrombin. The latter treatment totally removed the fibrinogen line.4. A non-fibrinogen precipitin line, observed only with the antiserum to platelet extract and positioned in the β2-globulin region, disappeared by the incubation of platelet extracts with thrombin.

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