STUDIES ON ϵ-PROTOTOXIN OF CLOSTRIDIUM PERFRINGENS TYPE D

1964 ◽  
Vol 42 (4) ◽  
pp. 545-554 ◽  
Author(s):  
A. F. S. A. Habeeb
Keyword(s):  
Clostridium Perfringens ◽  
Moving Boundary ◽  
Deae Cellulose ◽  
Paper Electrophoresis ◽  
Disc Electrophoresis ◽  
Type D ◽  
Sedimentation Constant ◽  
Diffusion Constants ◽  
Starch Gel ◽  
And Diffusion

Purified prototoxin of Clostridium perfringens type D was separated by column chromatography on CM-cellulose or DEAE-cellulose into two and three fractions respectively. The fractions exhibited prototoxin activity and showed immunochemical identity. The purified prototoxin gave a single component in the ultracentrifuge with a sedimentation constant of 2.85 S. The molecular weight was 25,100 ± 1500 and 23,200 when determined by the Archibald method and from sedimentation and diffusion constants respectively. Although the prototoxin was homogeneous by paper electrophoresis and moving boundary electrophoresis at pH 4.5, its heterogeneity was demonstrated on moving boundary electrophoresis at pH 8.6; 72% of material had a mobility of −1 × 10−5 cm2/v sec and 28% with a mobility of −2.7 × 10−5 cm2/v sec. The electrophoretic heterogeneity was also demonstrated on starch gel and disc electrophoresis. Amino acid analysis showed the presence of hydroxyproline and four uncommon amino acids; two of the latter were identified tentatively as hydroxylysine and allohydroxylysine.

A STUDY OF THE SOLUBLE ANTIGENS OF CLOSTRIDIUM PERFRINGENS TYPE D

1964 ◽  
Vol 10 (2) ◽  
pp. 287-298 ◽  
Author(s):  
A. F. S. A. Habeeb
Keyword(s):  
Biological Activity ◽  
Clostridium Perfringens ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Double Diffusion ◽  
Starch Gel Electrophoresis ◽  
Type D ◽  
Starch Gel

The soluble antigens of Clostridium perfringens type D have been separated into four fractions by chromatography on DEAE-cellulose. The fractions were examined for their biological activity and for their homogeneity by starch gel electrophoresis, double diffusion in agar, and immunoelectrophoresis.

The Apparent Microheterogeneous Nature of the High-Sulfur Proteins of α-Keratins

1969 ◽  
Vol 39 (3) ◽  
pp. 273-279 ◽  
Author(s):  
L. S. Swart ◽  
F. J. Joubert ◽  
A. J. C. Strydom
Keyword(s):  
Tryptic Peptide ◽  
Moving Boundary ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Disc Electrophoresis ◽  
Single Peak

The homogeneity of wool SCMK-B2 and mohair SCMK-B2, prepared according to the procedure of Gillespie, has been investigated. Moving boundary electrophoresis and gel filtration of the preparations revealed a single peak. Examination of the SCMK-B2 preparations by disc electrophoresis demonstrated the presence of various components. The preparations were each separated by chromatography on DEAE-cellulose into four subfractions. The aminoacid composition and tryptic peptide patterns of the subfractions were similar but certain characteristic differences were obvious.

scholarly journals THE MOLECULAR WEIGHT AND ISOELECTRIC POINT OF THYROGLOBULIN

1935 ◽  
Vol 19 (1) ◽  
pp. 95-108 ◽  
Author(s):  
Michael Heidelberger ◽  
Kai O. Pedersen
Keyword(s):  
Molecular Weight ◽  
Isoelectric Point ◽  
Specific Volume ◽  
Sedimentation Equilibrium ◽  
Sedimentation Constant ◽  
Diffusion Constants ◽  
Equilibrium Data ◽  
Round Numbers ◽  
And Diffusion

1. The sedimentation constant of hog thyroglobulin is 19.2ċ10–13. That of human thyroglobulin is essentially the same. 2. The specific volume of hog thyroglobulin is 0.72. 3. The isoelectric point of native hog thyroglobulin is at pH 4.58, that of denatured thyroglobulin at pH 5.0. 4. The molecular weight of hog thyroglobulin is, in round numbers, 700,000, as calculated from the sedimentation and diffusion constants, or 650,000, as calculated from the sedimentation equilibrium data. 5. The thyroglobulin molecule deviates markedly from the spherical.

scholarly journals Studies on Reduced Wool III. Starch-Gel Electrophoresis of Extracted Wool Proteins

1964 ◽  
Vol 17 (1) ◽  
pp. 277 ◽  
Author(s):  
EOP Thompson ◽  
IJ O'donnell
Keyword(s):  
Gel Electrophoresis ◽  
Moving Boundary ◽  
Electrophoretic Pattern ◽  
Deae Cellulose ◽  
Protein Fractions ◽  
Single Peak ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
Wool Proteins

Starch-gel electrophoresis in 8M urea has been used to demonstrate the presence of many components in wool protein fractions extracted from reduced and alkylated wool. All preparations of low-sulphur wool proteins gave mUltiple bands on starch gel in 8M urea even though some of these had previously been fractionated to give a single peak using moving-boundary electrophoresis in the absence of 8M urea. The heterogeneity suggested by these results is in Rccord with that found by chromatography of the proteins on DEAE-cellulose in buffers containing 8M urea. With stepwise elution from DEAE-cellulose it is possible to obtain fractions responsible for various sections of the starch-gel electrophoretic pattern.

scholarly journals THE PROTEINS IN UNHEATED CULTURE FILTRATES OF HUMAN TUBERCLE BACILLI

1948 ◽  
Vol 87 (3) ◽  
pp. 229-244 ◽  
Author(s):  
Ellen B. Bevilacqua ◽  
Janet R. McCarter
Keyword(s):  
Molecular Weight ◽  
Diffusion Rate ◽  
Sedimentation Velocity ◽  
Partial Specific Volume ◽  
Velocity Measurements ◽  
Physical Measurements ◽  
Sedimentation Constant ◽  
Diffusion Constants ◽  
Culture Filtrates ◽  
And Diffusion

Concentrated culture filtrates of two strains of human tubercle bacilli, a virulent and a slightly virulent one, have been fractionated to give fourteen fractions in each case. Chemical determinations and sedimentation velocity measurements have been carried out on those fractions for which significant results could be obtained. The evidence is that two distinct proteins are present, in addition to a polysaccharide and nucleic acid. The physical measurements have not demonstrated the presence of any other proteins. One of the proteins has been isolated in pure form, and found to have a molecular weight of 44,000 ± 5,000, based on measurements of partial specific volume, sedimentation velocity, and diffusion rate. This protein is believed to be the same as one previously isolated by Seibert et al. (6), who assigned it a molecular weight of 32,000. The other protein was not obtained sufficiently free from polysaccharide so that its molecular weight could be determined, but it is believed to have a sedimentation constant of about 2 S. Sedimentation and diffusion constants have been obtained for the polysaccharide, which appears to be a homogeneous molecular species with a molecular weight of about 20,000. The source in unheated tuberculin of the proteins obtained from heated preparations is discussed.

Bovine Platelet Proteins III. Some Properties of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 428-440 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Intrinsic Viscosity ◽  
Rabbit Antiserum ◽  
Disc Electrophoresis ◽  
Carbohydrate Content ◽  
Plasma Fibrinogen ◽  
Total Amino Acid ◽  
Electrophoretic Mobilities ◽  
Bovine Plasma ◽  
Starch Gel ◽  
The Difference

Summary1. Some properties of purified bovine platelet fibrinogen have been described and the data compared to those obtained by parallel analysis of purified bovine plasma fibrinogen.2. A close similarity was found between platelet and plasma fibrinogen as to sedimentation coefficients, electrophoretic mobilities in starch gel and polyacrylamide disc electrophoresis, light absorption spectra in the range 240 mμ to 330 mμ, ability to form immunoprecipitate with a rabbit antiserum against bovine plasma fibrinogen, total amino acid composition and in N-terminal amino acids.Differences between the fibrinogens were found as to intrinsic viscosity, carbohydrate content and behaviour upon clotting by thrombin. Intrinsic viscosity in 0.3 M NaCl at 25° was 0.48 dl/g for platelet fibrinogen as compared to 0.26 dl/g for plasma fibrinogen. The carbohydrate content of platelet fibrinogen was 0.56 ± 0.10% 1.56±0.10% and 1.37±0.09% for sialic acid (calculated as N-glycolyl neuraminic acid), hexose (galactose/mannose 1:2) and hexosamine (glucosamine), respectively. These values were 6, 54 and 26% higher than those found for plasma fibrinogen. The difference in clotting behaviour indicated a slower polymerization rate of the fibrin monomers formed from platelet fibrinogen than of those formed from plasma fibrinogen.

Bovine Platelet Proteins II. Purification of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 419-427 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Analytical Ultracentrifugation ◽  
Insoluble Fraction ◽  
Disc Electrophoresis ◽  
Plasma Fibrinogen ◽  
Starch Gel Electrophoresis ◽  
Freezing And Thawing ◽  
Insoluble Material ◽  
High Resolution Power ◽  
Starch Gel ◽  
Platelet Proteins

Summary1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 109 platelets. Total protein of this fraction was 0.77 ±0.08 mg per 109 platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 109 platelets.3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

Anticoagulants Produced by Thrombin from Fibrin, the Effect on Blood Coagulation, Some Physical Characteristics

1971 ◽  
Vol 26 (02) ◽  
pp. 211-223 ◽  
Author(s):  
Ch R. Muirhead ◽  
D. C Triantaphyllopoulos
Keyword(s):  
Blood Coagulation ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Fibrin Clot ◽  
Disc Electrophoresis ◽  
Physical Characteristics ◽  
Advanced Stage ◽  
Kallikrein Inhibitor ◽  
Shed Blood ◽  
Complete Lysis

SummaryChromatographed thrombin in the presence of both 50 Kallikrein inhibitor units of Trasylol per ml and 0.1 M E-ACA solubilized fibrin and the products of lysis possessed anticoagulant properties. The peak of the antithrombic activity coincided with the time of complete lysis of the fibrin clot, plasmin lysed fibrin exhibited the peak of its antithrombic activity much earlier. The effect of thrombin lysed fibrin on the prothrombin consumption of shed blood was found to be inhibitory.The products of the digestion of fibrin by thrombin and by plasmin, isolated at an advanced stage of proteolysis were compared by gel filtration, disc electrophoresis and DEAE cellulose chromatography. Differences in physical characteristics of these fibrin breakdown products offer evidence that they were produced by two different enzymes.

INHIBITION BY HUMAN SERUM OF THE ADIPOKINETIC EFFECT OF A HUMAN PITUITARY LIPID-MOBILIZING FACTOR

1972 ◽  
Vol 71 (3) ◽  
pp. 443-453 ◽  
Author(s):  
Olav Trygstad ◽  
Irene Foss
Keyword(s):  
Human Serum ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Inhibitory Effect ◽  
Disc Electrophoresis ◽  
Inhibitory Protein ◽  
Human Pituitary ◽  
Albumin Fraction ◽  
Pituitary Glands ◽  
Normal Human

ABSTRACT A lipid-mobilizing factor (LMF) with an adipotrophic effect in human and animal fat tissue has been prepared from human pituitary glands. The addition of normal human serum to LMF reduced its lipolytic effect, and it was completely abolished by serum from a group of obese patients, whereas the lipolysis was not influenced by serum from patients with generalized lipodystrophy. By DEAE-cellulose chromatography of human serum the inhibitory effect on LMF was found to be present in a protein fraction less acidic than the main serum albumin fraction. The inhibitory fraction was deprived of some contaminants by Sephadex gel filtration. Disc electrophoresis demonstrated the presence of three components in the inhibitory protein (IP), and they were identified as albumin, transferin, and haemopexin by immuno-electrophoresis. Precipitation of these proteins by their rabbit antisera demonstrated that the inhibitory effect was present in the albumin fraction. Insulin like activity was not observed in IP. A protein binding of LMF by IP could not be demonstrated. Incubation at 37°C for one hour of a mixture of LMF and IP eliminated the electrophoretic picture of LMF. It is concluded that the inhibitory effect of human serum may be due to proteolysis of LMF.

ULTRACENTRIFUGE AND DIFFUSION STUDIES ON GLUTEN

1942 ◽  
Vol 20c (3) ◽  
pp. 130-159 ◽  
Author(s):  
A. G. McCalla ◽  
Nils Gralén
Keyword(s):  
Sodium Salicylate ◽  
Minimum Weight ◽  
Average Molecular Weight ◽  
Sedimentation Equilibrium ◽  
Velocity Sedimentation ◽  
Molecular Characteristics ◽  
Weight Average Molecular Weight ◽  
Diffusion Constants ◽  
Diffusion Studies ◽  
And Diffusion

The molecular characteristics of gluten in sodium salicylate solutions were studied by means of sedimentation velocity, sedimentation equilibrium, and diffusion measurements. The proportion of total gluten protein molecularly dispersed increased with increase in concentration of sodium salicylate up to 12%, but the dispersed portions had essentially the same sedimentation constant (2.5 ± 0.15) regardless of the concentration of the dispersing medium.The most soluble 25 per cent of the gluten was all molecularly dispersed, but was definitely inhomogeneous. The weight-average molecular weight of this fraction was 44,000, but there is reason to believe the minimum weight may be about 35,000. None of the other fractions was entirely molecularly dispersed, the proportion decreasing with decreasing solubility of the fractions. Aggregates of many sizes existed in all of these fractions, but only the most insoluble contained aggregates large enough to cause opacity. Sedimentation constants of the molecularly dispersed portions increased slightly with decreasing solubility, while diffusion constants decreased markedly. None of the fractions yielded normal curves (diffusion diagrams) but the more soluble the fraction, the more nearly normal the curve. The inhomogeneity responsible for the varying rates of diffusion was due partly to differences in proportion and properties of the molecularly dispersed gluten and partly to aggregates.All properties showed progressive changes both within and between the arbitrarily produced fractions. These results, therefore, support the hypothesis that gluten is a protein system showing progressive and regular changes in properties with change in solubility.

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