An electronic colloid osmometer and an assessment of its accuracy. The molecular weight of bovine plasma albumin

The molecular weight of performic acid oxidized bovine plasma albumin, dispersed in 0.08 M borate +0.2 M sodium chloride buffer, pH 7.4, was estimated as 30,000 by light-scattering and sedimentation equilibrium methods, 19,000 by osmotic pressure. Sedimentation velocity analyses and electrophoresis showed that the component polypeptide chains of the material are similar in mass and charge density so the polydispersity must be attributed to labile aggregates. The results indicate that here are at least three and probably four similar polypeptide chains in the molecule of native bovine plasma albumin, held together by disulphide bonds.

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PURIFICATION AND PROPERTIES OF A PROTEASE FROM PENICILLIUM CYANEO-FULVUM

Canadian Journal of Biochemistry and Physiology ◽

10.1139/y60-121 ◽

1960 ◽

Vol 38 (1) ◽

pp. 969-980 ◽

Cited By ~ 2

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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MOLECULAR WEIGHT OF BOVINE PLASMA ALBUMIN, VITELLENIN, AND THEIR OXIDATION PRODUCTS IN FORMIC ACID

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o61-150 ◽

1961 ◽

Vol 39 (9) ◽

pp. 1405-1417 ◽

Cited By ~ 6

Author(s):

W. G. Martin

Keyword(s):

Molecular Weight ◽

Formic Acid ◽

Transient State ◽

Oxidation Products ◽

Performic Acid ◽

Bond Rupture ◽

Acid Oxidation ◽

Plasma Albumin ◽

Viscosity Measurements ◽

Bovine Plasma

Sedimentation, diffusion, and Archibald transient state measurements were made on bovine plasma albumin and vitellenin of egg yolk in formic acid (88% w/w) solution. The molecular weight of bovine plasma albumin, averaging 77 × 103and 70 × 103with and without added salt, respectively, indicated that peptide bonds were stable to the acid for at least 1 week (storage at 5 °C and measurement periods at 20 °C). Similar values were obtained from estimates based on viscosity measurements but greater deviations occurred. Vitellenin had a mean molecular weight of 93 × 103from sedimentation and diffusion but polydispersity was revealed by the Archibald measurements (molecular weights from 55 × 103to 10 × 103). Higher values of molecular weight were obtained for vitellenin by varying the dissolution technique and exposure time in formic acid and also when viscosity measurements were used to compute molecular weight. Analyses of N-terminal amino acids showed that peptide bond rupture was not a major factor in the polydispersity of vitellenin. Although aggregates are probably present in formic acid solutions of this protein, it appears to be naturally polydisperse. Both albumin and vitellenin were considerably degraded by performic acid oxidation procedures.

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DEGRADATION BY HYDRAZINE OF BOVINE PLASMA ALBUMIN AND LYSOZYME

Canadian Journal of Biochemistry and Physiology ◽

10.1139/y60-089 ◽

1960 ◽

Vol 38 (1) ◽

pp. 715-726 ◽

Cited By ~ 1

Author(s):

Madeleine Champagne ◽

David B. Smith

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Hydrazine Hydrate ◽

Plasma Albumin ◽

Bovine Plasma ◽

Terminal Amino

The effect of hydrazine and hydrazine hydrate on bovine plasma albumin and lysozyme at 5° and 20 °C has been studied by viscosity, sedimentation, and molecular weight measurements. The appearance of new N-terminal amino acids and peptides has been demonstrated. The effect of these reagents is an initial unfolding of the molecule followed by slow, non-random fission to smaller particles.

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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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DEGRADATION BY HYDRAZINE OF BOVINE PLASMA ALBUMIN AND LYSOZYME

Canadian Journal of Biochemistry and Physiology ◽

10.1139/o60-089 ◽

1960 ◽

Vol 38 (7) ◽

pp. 715-726 ◽

Cited By ~ 4

Author(s):

Madeleine Champagne ◽

David B. Smith

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Hydrazine Hydrate ◽

Plasma Albumin ◽

Bovine Plasma ◽

Terminal Amino

The effect of hydrazine and hydrazine hydrate on bovine plasma albumin and lysozyme at 5° and 20 °C has been studied by viscosity, sedimentation, and molecular weight measurements. The appearance of new N-terminal amino acids and peptides has been demonstrated. The effect of these reagents is an initial unfolding of the molecule followed by slow, non-random fission to smaller particles.

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The isolotion of serologically active peptides of low molecular weight from a hydrolyzate of bovine plasma albumin

Archives of Biochemistry and Biophysics ◽

10.1016/0003-9861(60)90584-1 ◽

1960 ◽

Vol 90 (2) ◽

pp. 309-313 ◽

Cited By ~ 7

Author(s):

Alfred J. Richard ◽

Sherwin Beck ◽

Hans Hoch

Keyword(s):

Molecular Weight ◽

Low Molecular Weight ◽

Plasma Albumin ◽

Active Peptides ◽

Bovine Plasma

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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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