STUDIES ON PLASMINOGEN: VI. ACTIVATION PRODUCTS OF BOVINE AND HUMAN PLASMINOGENS

1966 ◽  
Vol 44 (4) ◽  
pp. 487-495 ◽  
Author(s):  
John Y. S. Chan ◽  
Edwin T. Mertz
Keyword(s):  
Gel Electrophoresis ◽  
Elution Curve ◽  
Major Protein ◽  
Starch Gel Electrophoresis ◽  
Sephadex Column ◽  
Molecular Weights ◽  
Activation Products ◽  
Starch Gel ◽  
Human Plasminogen

Activated, purified bovine plasminogen, when fractionated on a Sephadex column, revealed four major protein peaks. Bovine plasmin I and plasmin II were found under peak II and peak III, respectively. Highly potent bovine plasmin II was recovered from peak III. Activated human plasminogen, when fractionated by the same technique, yielded an elution curve similar to that of the nonactivated sample. The pathway of the breakdown of bovine plasminogen upon activation with urokinase was studied by starch-gel electrophoresis and Sephadex chromatography. It is suggested that native bovine plasminogen is first converted to an altered plasminogen, which is then converted to plasmin I, plasmin I is then converted to plasmin II, and plasmin II to peptide A. The molecular weights of plasmin I, plasmin II, and peptide A were estimated by the Sephadex method to be 8.0 × 104, 6.2 × 104, and 4.3 × 104, respectively.

STUDIES ON PLASMINOGEN: V. PURIFICATION OF BOVINE AND HUMAN PLASMINOGENS BY SEPHADEX CHROMATOGRAPHY

1966 ◽  
Vol 44 (4) ◽  
pp. 475-485 ◽  
Author(s):  
John Y. S. Chan ◽  
Edwin T. Mertz
Keyword(s):  
Human Serum ◽  
Column Chromatography ◽  
Continuous Flow ◽  
Sephadex Column ◽  
Molecular Weights ◽  
Starch Gel ◽  
Electrophoresis Method ◽  
Human Plasminogen ◽  
Specific Activities

Purified euglobulins prepared from bovine and human serum were fractionated by Sephadex column chromatography at pH 3.5. The products obtained were purified further by phosphate precipitation (bovine) or lysine solubilization (human) to yield final products with activities comparable to those prepared by the continuous-flow electrophoresis method. Plasminogen from Cohn fraction III-4 was also purified. The method yields preparations from serum which contain essentially no altered plasminogen. The specific activities, the starch-gel patterns, the molecular weights, and the recoveries of the purified bovine and human plasminogen preparations are reported and discussed.

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.

scholarly journals Some relationships between R-factor and chromosomal β-lactamase in Gram-negative bacteria

1971 ◽  
Vol 123 (4) ◽  
pp. 507-512 ◽  
Author(s):  
J. W. Dale ◽  
J. T. Smith
Keyword(s):  
Gel Electrophoresis ◽  
Klebsiella Aerogenes ◽  
Gram Negative Bacteria ◽  
Starch Gel Electrophoresis ◽  
Gram Negative ◽  
E Coli ◽  
Molecular Weights ◽  
R Factor ◽  
Starch Gel ◽  
R Factors

1. The β-lactamases specified by Klebsiella aerogenes 418 and the R-factor R-7268 have been partially purified. 2. The molecular weights of the K. aerogenes strains 418 and 373, Aerobacter cloacae 53, R-7268 and R-TEM β-lactamases were all about 20000; that of the enzymes from Escherichia coli strains 419 and 214T was about 31000. 3. These enzymes were also compared by means of their Km values for benzylpenicillin and ampicillin, and their behaviour on starch-gel electrophoresis. 4. The β-lactamases specified by the two Klebsiella strains, the Aerobacter strain, and the R-factors R-TEM and R-7268 were found to comprise a broadly similar group. However, within this group, only two enzymes seemed to be identical, namely those specified by the two R-factors. The two E. coli strains produce identical β-lactamases which are very different from the ‘Klebsiella/Aerobacter-type’ enzymes.

The labelling of certain milk proteins with iodine131

1965 ◽  
Vol 32 (2) ◽  
pp. 181-186 ◽  
Author(s):  
H. A. Veringa ◽  
M. F. Kerkhof Mogot
Keyword(s):  
Gel Electrophoresis ◽  
Milk Proteins ◽  
Starch Gel Electrophoresis ◽  
Molecular Weights ◽  
Fat Globules ◽  
Starch Gel ◽  
Clustering Effect ◽  
Β Lactoglobulin ◽  
Electrophoresis Pattern

SummaryWhole casein, β-casein, β-lactoglobulin and euglobulin were labelled with 131I. The conditions under which iodination was carried out were chosen so as to avoid any modification of the original characteristics of the proteins. This was checked by starch-gel electrophoresis and determination of sedimentation constants, apparent molecular weights and, for euglobulin, the clustering effect on fat globules.As was shown by autoradiograms of the starch-gel plates, the radioactivity was incorporated in all zones of the electrophoresis pattern.

scholarly journals Electrophoretic and immunological charactorization of rat neurophysin

1971 ◽  
Vol 122 (5) ◽  
pp. 671-676 ◽  
Author(s):  
A. Norström ◽  
J. Sjöstrand ◽  
B. G. Livett ◽  
L. O. Uttenthal ◽  
D. B. Hope
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Minor Component ◽  
Polyacrylamide Gel ◽  
Male Rats ◽  
Major Protein ◽  
Starch Gel Electrophoresis ◽  
Neural Lobe ◽  
Starch Gel

1. The electrophoretic properties of rat posterior pituitary proteins have been compared on starch gel with those of bovine and porcine neurophysins. 2. [35S]-Cysteine was injected into the supraoptic nucleus of male rats and 16–24h later the distribution of labelled neural-lobe protein in starch and polyacrylamide gels was determined. In both systems a single major protein component was found to contain more than 80% of the total recovered radioactivity. Between 5 and 10% of the radioactivity was found in a minor component in polyacrylamide gel. 3. In agar, microimmuno-diffusion and -electrophoresis of the rat neural-lobe proteins gave a single arc with neurophysin antiserum, and after starch-gel electrophoresis this arc was shown to be due to the major labelled component. 4. The molecular weights of the rat neural-lobe proteins were estimated by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The molecular weight of the major labelled component was found to be 12000. 5. It is concluded that the rat neurophysin consists of one major and possibly one minor component.

SPECIES, TISSUE, AND INDIVIDUAL SPECIFICITY OF LOW IONIC STRENGTH EXTRACTS OF AVIAN MUSCLE AND OTHER ORGANS REVEALED BY STARCH-GEL ELECTROPHORESIS

1966 ◽  
Vol 44 (6) ◽  
pp. 853-859 ◽  
Author(s):  
C. M. Ann Baker
Keyword(s):  
Ionic Strength ◽  
Gel Electrophoresis ◽  
Species Specificity ◽  
Major Protein ◽  
Starch Gel Electrophoresis ◽  
Avian Muscle ◽  
Starch Gel ◽  
Low Ionic Strength ◽  
Individual Specificity ◽  
Electrophoretic Resolution

The proteins soluble at low ionic strength of various muscles and of other tissues from five species of birds were examined by vertical starch-gel electrophoresis. The methods used were simple, and gave excellent and repeatable electrophoretic resolution of proteins. Most samples yielded 15–25 zones which stained nonspecifically for protein. Histochemical techniques revealed additional, enzyme, bands which were not coincident with the "major" protein zones. The results confirm and extend previous observations of the species specificity of the electrophoretic profiles of proteins from muscle extracts (myogen), and reveal considerable tissue and individual specificity of the enzymes and other proteins in extracts of avian tissues.

scholarly journals Die Trennung der Hämolymphe-Proteine bei Vorpuppen und Puppen der Galleria mellonella L. mittels Gel-Filtration auf Sephadex G-200, Bestimmung ihrer isoelektrischen Punkte und ihrer Molekulargewichte / Separation of Haemolymph Proteins of Prepuae and Pupae of Galleria mellonella L. by means of Gel Filtration on Sephadex 200, Determination of their Isoelectric Points and their Molecular Weights

1969 ◽  
Vol 24 (6) ◽  
pp. 732-740 ◽  
Author(s):  
Milan Marek
Keyword(s):  
Gel Electrophoresis ◽  
Galleria Mellonella ◽  
Gel Filtration ◽  
Starch Gel Electrophoresis ◽  
Molecular Weights ◽  
Isoelectric Points ◽  
Starch Gel ◽  
The Individual ◽  
Naphthyl Acetate

Gel filtration on Sephadex G-200 was carried out on haemolymph proteins of prepupae. ligatured prepupae, male and female pupae and cooled pupae of Galleria mellonella L.The proteins were separated into two main fractions. The esterase activity of the eluated haemolymph was determined by means of beta-naphthyl acetate after filtration.After elution the samples were condensed and additionally separated on horizontal starch-gel electrophoresis.The “cooling protein” of pupae and the “ligature protein” of ligatured larvae of Galleria mellonella were shown by means of starch-gel electrophoresis to be new proteins, so far not described.The isoelectric point and molecular weight were determined for the individual protein fractions.They were then stained with amido black 10 B for the proof of proteins, and with alpha-naphthyl butyrate with Fast-Blue BB salt for the identification of esterases.

STUDIES ON PLASMINOGEN: III. PURIFICATION OF BOVINE AND HUMAN PLASMINOGEN BY CONTINUOUS ELECTROPHORESIS

1963 ◽  
Vol 41 (8) ◽  
pp. 1811-1819 ◽  
Author(s):  
Edwin T. Mertz ◽  
John Y. S. Chan
Keyword(s):  
Human Blood ◽  
Gel Electrophoresis ◽  
Cathode Side ◽  
Human Blood Plasma ◽  
Starch Gel Electrophoresis ◽  
Anode Side ◽  
Cold Room ◽  
Bovine Plasma ◽  
Starch Gel ◽  
Human Plasminogen

Euglobulin from bovine plasma or outdated human blood plasma was freed of interfering impurities and applied to the anode side of the paper curtain of a Beckman Spinco CP electrophoresis apparatus in a cold room. Background buffer was at pH 3.5. The first and second leading fractions (cathode side) were combined and they contained plasminogen with approximately 10 times the specific esterolytic and caseinolytic activities of the original euglobulin. Cohn Fraction III can be substituted for human euglobulin. Additional purification can be obtained by phosphate precipitation of the curtain products. The specific esterolytic and caseinolytic activities of purified bovine and human plasminogen preparations, as well as their starch gel electrophoresis patterns, are reported and discussed.

STUDIES ON PLASMINOGEN: III. PURIFICATION OF BOVINE AND HUMAN PLASMINOGEN BY CONTINUOUS ELECTROPHORESIS

1963 ◽  
Vol 41 (1) ◽  
pp. 1811-1819
Author(s):  
Edwin T. Mertz ◽  
John Y. S. Chan
Keyword(s):  
Human Blood ◽  
Gel Electrophoresis ◽  
Cathode Side ◽  
Human Blood Plasma ◽  
Starch Gel Electrophoresis ◽  
Anode Side ◽  
Cold Room ◽  
Bovine Plasma ◽  
Starch Gel ◽  
Human Plasminogen

Euglobulin from bovine plasma or outdated human blood plasma was freed of interfering impurities and applied to the anode side of the paper curtain of a Beckman Spinco CP electrophoresis apparatus in a cold room. Background buffer was at pH 3.5. The first and second leading fractions (cathode side) were combined and they contained plasminogen with approximately 10 times the specific esterolytic and caseinolytic activities of the original euglobulin. Cohn Fraction III can be substituted for human euglobulin. Additional purification can be obtained by phosphate precipitation of the curtain products. The specific esterolytic and caseinolytic activities of purified bovine and human plasminogen preparations, as well as their starch gel electrophoresis patterns, are reported and discussed.

scholarly journals STUDIES ON STRUCTURAL UNITS OF THE γ-GLOBULINS

1961 ◽  
Vol 113 (5) ◽  
pp. 861-884 ◽  
Author(s):  
G. M. Edelman ◽  
M. D. Poulik
Keyword(s):  
Amino Acid ◽  
Gel Electrophoresis ◽  
Disulfide Bonds ◽  
Average Molecular Weight ◽  
Structural Features ◽  
Starch Gel Electrophoresis ◽  
Molecular Weights ◽  
Γ Globulins ◽  
Starch Gel ◽  
Polypeptide Chains

When human and rabbit 7S γ-globulins were reduced in strong urea solutions by a number of procedures, their molecular weights fell to approximately ⅓ of the original values. Partial separation of the reduction products was achieved using chromatography and starch gel electrophoresis in urea solutions. One of the components of reduced human 7S γ-globulin was isolated by chromatography, identified by starch gel electrophoresis, and subjected to amino acid analyses. The amino acid composition of this component differed from that of the starting material and also from that of the remaining components. A reduced pathological macroglobulin dissociated to components with an average molecular weight of 41,000. Several reduced human myeloma proteins, when subjected to starch gel electrophoresis, yielded individual patterns that nevertheless had features in common with those of reduced normal γ-globulins. Reduction of normal and abnormal γ-globulins was accompanied by the appearance of titratable sulfhydryl groups. Chemical treatments other than reduction were used to determine the type of bond holding the subunits together. It was tentatively concluded that they were linked by disulfide bonds. An hypothesis is presented to relate the structural features of the various γ-globulins in terms of the multiplicity of polypeptide chains in these molecules.

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