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.

scholarly journals A chromatographic preparation of ox growth hormone

1966 ◽  
Vol 100 (3) ◽  
pp. 593-600 ◽  
Author(s):  
M Wallis ◽  
HBF Dixon
Keyword(s):  
Molecular Weight ◽  
Growth Hormone ◽  
Gel Electrophoresis ◽  
Anterior Pituitary ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Highly Active ◽  
Cross Linkage ◽  
Starch Gel

1. A method is described for the chromatographic preparation of ox growth hormone. It involves chromatography of an extract of anterior pituitary lobes on DEAE-cellulose, followed by rechromatography on a dextran gel of low cross-linkage (Sephadex G-100). 2. The product is highly active in growth-hormone assays, and is obtained in good yield. It was homogeneous by several criteria, but showed some heterogeneity on starch-gel electrophoresis. 3. The molecular weight of the hormone was estimated from its behaviour on gel-filtration columns under various conditions. Evidence that the hormone may dissociate into sub-units under some conditions is presented.

STUDIES ON PLASMINOGEN: IV. ALTERATION OF BOVINE AND HUMAN PLASMINOGENS DURING ISOLATION

1966 ◽  
Vol 44 (4) ◽  
pp. 469-473 ◽  
Author(s):  
John Y. S. Chan ◽  
Edwin T. Mertz
Keyword(s):  
Ionic Strength ◽  
Gel Electrophoresis ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Major Band ◽  
Starch Gel ◽  
Human Plasminogen

Bovine and human plasminogen preparations were analyzed by starch-gel electrophoresis at pH 2.5 and 0.10 ionic strength. The bands were activated with urokinase and the proteolytic and esterolytic activities measured. Bovine euglobulin contains one plasminogen band, B-1. Plasminogen prepared from bovine euglobulin by continuous electrophoresis at pH 3.5 contains B-1 and a faster plasminogen band, B-2. B-1 and B-2 are also found in bovine plasminogen prepared by DEAE-cellulose chromatography. All three preparations on activation give the same two plasmin bands on starch gel. Human euglobulin also contains two active plasminogen bands H-1 and H-2. Plasminogen prepared from human euglobulin by continuous electrophoresis at pH 3.5 contains H-1, H-2, and a faster minor plasminogen band, H-3. All highly purified human plasminogens derived from Cohn fraction III contain either H-3 as a major band and an additional plasminogen band, H-4, or only H-3, but no H-1 and H-2. On activation with urokinase or streptokinase, human plasminogen preparations give one or two plasmin bands. It is concluded that bovine B-2 and human H-3 and H-4 are altered forms of euglobulin plasminogen created during isolation procedures. Essentially pure human H-3 can be prepared by continuous electrophoresis from Cutter plasminogen.

Benzylamine oxidase and histaminase: purification and crystallization of an enzyme from pig plasma

1964 ◽  
Vol 161 (983) ◽  
pp. 153-167 ◽  
Keyword(s):  
Ammonium Sulphate ◽  
Gel Electrophoresis ◽  
Copper Content ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Anaerobic Conditions ◽  
Concentrated Solutions ◽  
Radioactivation Analysis ◽  
Starch Gel ◽  
Benzylamine Oxidase

The enzyme benzylamine oxidase of pig plasma has been purified and some of the properties of the pure preparation have been studied. The purification procedure included several precipitations with ammonium sulphate and separations of proteins by column chromatography, first on DEAE-cellulose, followed by DEAE-Sephadex and lastly on a hydroxyapatite column. Crystals were prepared from solutions of the purified enzyme by adding ammonium sulphate. The crystalline preparation was homogeneous when studied by starch-gel electrophoresis and by ultracentrifugation. The molecular weight, as determined on the analytical ultracentrifuge, was 195 000. The copper content of the enzyme, as determined by radioactivation analysis, was about four atoms of Cu per molecule of enzyme. Concentrated solutions of the enzyme had a pink colour; the colour disappeared when substrate (benzylamine) was added under anaerobic conditions. The amines which were tested and found to be oxidized by the pure enzyme were: benzylamine, histamine, mescaline and 4-picolylamine. The affinity of the enzyme for benzylamine was more than one hundred times that for histamine.

Studies on the relation between plasma antithrombin and heparin-cofactor

1968 ◽  
Vol 46 (3) ◽  
pp. 347-350 ◽  
Author(s):  
F. C. Monkhouse ◽  
Susan Milojevic
Keyword(s):  
Gel Electrophoresis ◽  
Specific Activity ◽  
Deae Cellulose ◽  
Fold Increase ◽  
Significant Loss ◽  
Starch Gel Electrophoresis ◽  
Electrophoretic Patterns ◽  
Starch Gel ◽  
Cofactor Activity ◽  
Cellulose Column

A method for the preparation of purified plasma antithrombin and heparin-cofactor is described. The method involves adsorption by aluminium hydroxide, separation on a DEAE-cellulose column by means of a graded salt concentration, and vertical curtain electrophoresis. A 100-fold increase in the specific activity of antithrombin and a 30-fold increase in the specific activity of heparin-cofactor have been achieved. In spite of the increased purification, no separation of the two activities was achieved. When a highly purified fraction was subjected to starch-gel electrophoresis for 16–18 h and then eluted from the gel, there was significant loss of heparin-cofactor activity but not of antithrombin activity. The electrophoretic patterns of the recovered proteins were not altered.

scholarly journals The separation and characterization of marmoset kidney β-d-galactosidase and β-d-glucosidase

1977 ◽  
Vol 167 (3) ◽  
pp. 765-773 ◽  
Author(s):  
R J Pierce ◽  
R G Price
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Apparent Molecular Weight ◽  
Exchange Chromatography ◽  
Starch Gel Electrophoresis ◽  
Lysosomal Fraction ◽  
Glucosidase Activity ◽  
Starch Gel

beta-D-Galactosidase and beta-D-glucosidase activities were determined in homogenates of marmoset kidney by using the appropriate 4-methylumbelliferyl glycoside, beta-D-Galactosidase activity was separated into two main components by ion-exchange chromatography on DEAE-cellulose, starch-gel electrophoresis, isoelectric focusing and gel filtration on Sephadex G-200. One form designated A had a pI of 5.1, was loosely bound to DEAE-cellulose at pH7.0, remained near the origin on starch-gel electrophoresis at pH 7.0 and had an apparent molecular weight of 160000. The second beta-D-galactosidase component, designated B, was associated with the total beta-D-glucosidase activity, had a pI of 4.3, was firmly bound to DEAE-cellulose, migrated rapidly towards the anode on starch-gel electrophoresis and had an apparent molecular weight of 50000. The optimum pH values of beta-D-galactosidase A and B were 4.5 and 6.0 respectively. beta-D-Galactosidase A was activated by 0.1 M-NaC1 but the activity of the B form was inhibited by 1 M-NaC1 at pH 4.5. beta-D-galactosidase had a bimodal distribution, the A form being recovered in the lysosomal fraction whereas the B form was present in the soluble fraction, as was the major portion of the beta-D-glucosidase activity. The lysosomal and soluble forms were further characterized by DEAE-cellulose chromatography.

Preparation and properties of β-casein from buffalo's milk

1975 ◽  
Vol 42 (1) ◽  
pp. 163-167 ◽  
Author(s):  
M. H. Abd El-Salam ◽  
Safinaz El-Shibiny
Keyword(s):  
Gel Electrophoresis ◽  
Tryptic Peptide ◽  
Polypeptide Chain ◽  
Deae Cellulose ◽  
Starch Gel Electrophoresis ◽  
Terminal Sequence ◽  
Single Polypeptide Chain ◽  
End Groups ◽  
Starch Gel ◽  
Single Polypeptide

Summaryβ-Casein from individual buffalo's milk was found to be homogeneous by starch-gel electrophoresis. β-Casein was separated from buffalo's milk by the method of Warner (1944) and purified by DEAE-cellulose chromatography.Buffalo β-casein possesses identical end-groups to those of cow β-casein; namely N-terminal arginine and assuming a single polypeptide chain a possible C-terminal sequence of Ile-Ile-Val. However, the amino-acid composition and the tryptic peptide patterns of the 2 proteins are not the same.

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.

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.

Isolation of κ-casein-like fractions from human whole casein by chromatography on thiol-Sepharose, hydroxyapatite and DEAE-cellulose

1981 ◽  
Vol 48 (3) ◽  
pp. 429-436 ◽  
Author(s):  
Jean-Marc Chobert ◽  
Francesco Addeo ◽  
Bruno Ribadeau Dumas
Keyword(s):  
Affinity Chromatography ◽  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Phosphate Buffer ◽  
Deae Cellulose ◽  
The Other ◽  
Starch Gel Electrophoresis ◽  
Sh Groups ◽  
Starch Gel

SummaryDifferent fractions of human κ-casein characterized by their sensitivity to chymosin were separated, after reduction, by affinity chromatography on thiol-Sepharose in a 0·1 m-Tris-HCl buffer, pH 7·0, containing 7 M-urea, 0·3 m-NaCl and 1 mm-EDTA. The fraction retained on the column and bearing the SH-groups was then chromatographed on hydroxyapatite in a 5 mm-phosphate buffer, pH 6·8, containing 0·2 m-KC1, 4·5 m-urea and 2 mm 2-mercaptoethanol. The fraction not retained on this column was purified by chromatography on DEAE-cellulose in a 6·6 m-urea, 0·02 m-imidazole, 0·02 m 2-mercaptoethanol buffer, pH 7·0. Two chymosinsensitive fractions were identified by starch-gel electrophoresis at pH 8·6; one of low electrophoretic mobility, the other a smeared band covering the whole distance of migration, in which several bands with mobilities greater than those of β-caseins could be detected.

scholarly journals N-acetyl-β-d-glucosaminidase in marmoset kidney, serum and urine

1978 ◽  
Vol 175 (3) ◽  
pp. 859-867 ◽  
Author(s):  
R J Pierce ◽  
R G Price ◽  
J S L Fowler
Keyword(s):  
Gel Electrophoresis ◽  
Human Liver ◽  
Deae Cellulose ◽  
Heat Stability ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Partial Purification ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
Serum And Urine

N-Acetyl-beta-D-glucosaminidase activities were determined in homogenates of marmoset kidney, in serum and in urine by using the 4-methylumbelliferyl substrate. The enzyme activity was separated into several components by DEAE-cellulose ion-exchange chromatography, starch-gel electrophoresis and isoelectric focusing. The kidney contained two major forms of the enzyme, A and B, which had similar pH optima and Km values. The A-form bound to DEAE-cellulose at pH 6.8, migrated towards the anode on starch-gel electrophoresis and had a pI of 5.0. The B-form did not bind to DEAE-cellulose at pH 6.8, remained near the origin on starch-gel electrophoresis and had a pI of 7.64. The isoenzymes also differed in heat stability, the B-form being the more stable. Serum contained B-form activity and, in addition, two intermediate forms (I1 and I2) were loosely bound to DEAE-cellulose. The serum A-form activity was less firmly bound to DEAE-cellulose than was the tissue A-form and was designated As. Serum from a pregnant marmoset contained a form which may be analogous to the human P-isoenzyme. Urine contained only a small amount of B-form activity, the majority being present in the A-form. The kidney A- and B-forms both had mol.wts. of 96000–100000 and the activity was predominantly lysosomal. Partial purification of the kidney A isoenzyme was undertaken. Immunoprecipitation studies indicated a relationship between marmoset kidney A-form and human liver A-form activity.

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