scholarly journals Physicochemical and Biochemical Properties of Commercially Available Urokinase Preparations

1977 ◽  
Author(s):  
M. Nishida ◽  
H. Nishimaki ◽  
S. Miyake ◽  
T. Suyama ◽  
S. Morisue
Keyword(s):  
Gel Electrophoresis ◽  
Molecular Form ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Biochemical Properties ◽  
Molecular Forms ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
The Difference ◽  
Fresh Urine

Comparative studies were made on eleven urokinase preparations commercially available. Analysis by both gel filtration and sodium dodecyl gel electrophoresis revealed the variety of molecular forms of the activator in the preparations showing molecular weight approximately 54,000 (A), 47,000 (B) and 34,000 (C). Starch gel electrophoresis at pH 8.8 indicated that (B) and (C) moved to anode whereas (A) and the active component of fresh urine slightly moved to the cathod. Proteolytic digestion of (A) produced the same component as shown by (B) and (C) on starch gel electrophoresis. Plasminogen activating activity of (B) and (C) was found to be less than that of (A) when measured by the procedure of CTA fibrinolytic method with the physiological blood level of plasminogen. The present data suggest that variety of molecular form in the preparation may be due to the difference of purification procedure in view of proteolytic degradation of the enzyme, and (A) seems to be the naturally occuring type of urokinase in urine.

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.

Comparison of the rates of proteolysis during ripening of Cheddar cheeses made with calf rennet and swine pepsin as coagulants

1974 ◽  
Vol 41 (2) ◽  
pp. 269-282 ◽  
Author(s):  
Margaret L. Green ◽  
P. M. D. Foster
Keyword(s):  
Gel Electrophoresis ◽  
Gluconic Acid ◽  
Gel Filtration ◽  
Total Activity ◽  
Starch Gel Electrophoresis ◽  
Protein Breakdown ◽  
Rate Of Development ◽  
Slow Decline ◽  
Starch Gel ◽  
Acid Lactone

SummaryThe rates of proteolysis during ripening were followed in cheeses made with either calf rennet or swine pepsin and either starter or δ-gluconic acid lactone (GAL) as a replacement for the starter. A gel-filtration column technique and starch-gel electrophoresis were used for analysis, and bacterial counts were made on all samples. Proteolysis was faster in cheeses made using GAL than in those made using starter and also slightly faster in GAL cheeses made with swine pepsin than in those made with rennet. Further, it was considerably slower in starter-containing cheeses made with swine pepsin than in those made with rennet. It is suggested that these differences were due to the much greater rate of development of acidity in cheeses made with GAL than in those made with starter, which resulted in more of the coagulant being incorporated into the curd in an active state. The rate of proteolysis in starter-containing cheeses appeared to follow a characteristic course, being initially slow, then markedly increasing with a later slow decline. It is suggested that the increase in the rate of proteolysis was due to an increase in the total activity of bacterial proteinases released by lysis of the bacteria. Indications were obtained that the coagulants and bacterial proteinases catalysed broadly similar patterns of protein breakdown in cheese, and that medium-sized peptides (mol. wt 9000–14000) were formed as definite intermediates in the process. The results also showed that rennet and swine pepsin remained active for at least 7 months in GAL cheeses, that rennet contributed significantly to proteolysis in starter-containing cheeses, and that swine pepsin was at least extensively inactivated and possibly completely inactivated during cheese-making with starter.

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.

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.

STUDIES ON HUMAN CHORIONIC GONADOTROPHIN

1968 ◽  
Vol 59 (1) ◽  
pp. 89-104 ◽  
Author(s):  
H. van Hell ◽  
R. Matthijsen ◽  
J. D. H. Homan
Keyword(s):  
Sialic Acid ◽  
Calcium Ions ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Starch Gel Electrophoresis ◽  
Sialic Acid Content ◽  
Starch Gel ◽  
Biological Potency

ABSTRACT Highly purified preparations of Human Chorionic Gonadotrophin (HCG), with potencies up to 18 800 IU/mg, were obtained from material containing about 1500 IU/mg, by using a fractionation with ethanol, CMSephadex chromatography and gel filtration with Sephadex G-100. Starch gel electrophoresis revealed that the most potent preparation still contained three components, while certain other fractions with much lower potencies were found to be homogeneous. Evidence is presented for the presence of various HCG components differing from each other in biological potency, electrophoretic mobility and sialic acid content. Within this material, components with higher electroporetic mobilities and higher sialic acid contents showed higher biological potencies. The molecular weight estimated in the absence of calcium ions was 62 000 ± 1000.

scholarly journals The nature of the multiple forms of cytoplasmic aspartate aminotransferase from pig and sheep heart

1974 ◽  
Vol 141 (2) ◽  
pp. 401-406 ◽  
Author(s):  
Robert John ◽  
Richard Jones
Keyword(s):  
Aspartate Aminotransferase ◽  
Gel Electrophoresis ◽  
Starch Gel Electrophoresis ◽  
Wide Range ◽  
Covalent Structure ◽  
Starch Gel ◽  
The Difference ◽  
Ph Range ◽  
The Relationship ◽  
Charged Group

Starch-gel electrophoresis of sheep heart aspartate aminotransferase was carried out over the range pH7.0–8.5. The enzyme separates into three subforms in the same way as the pig heart enzyme. As the pH was increased the distance migrated by each subform increased by the same amount, so that they remained the same distance apart. Titration of the enzyme over the appropriate pH range was used to calculate the difference in charge between the subforms and it was concluded that they differ by one charged group per dimer from their nearest neighbour on the electrophoretogram over the whole pH range studied. It was also shown that the pig-heart α and β subforms differ by almost one charged group per dimer in the range pH5.5–5.7 and that the spacing between the subforms on starch-gel electrophoresis at pH8.0 is the same as that for the sheep enzyme. Since the charge difference between the subforms is maintained over such a wide range of pH, it is concluded that they probably differ from each other in covalent structure, because of the improbability that conformational differences can give rise to such behaviour. The relationship between the subforms and inactive binding of the coenzyme is also examined.

ASSOCIATION OF SERUM HEMOPEXIN, TRANSFERRIN, PREALBUMIN AND ALBUMIN1 TYPES WITH PRODUCTIVE TRAITS OF YORKSHIRE AND LACOMBE BREEDS OF PIGS

1969 ◽  
Vol 49 (2) ◽  
pp. 223-229 ◽  
Author(s):  
V. N. Tripathi ◽  
W. E. Howell
Keyword(s):  
Serum Protein ◽  
Gel Electrophoresis ◽  
Performance Test ◽  
Carcass Characteristics ◽  
Correlated Response ◽  
Starch Gel Electrophoresis ◽  
Starch Gel ◽  
The Difference ◽  
Productive Traits ◽  
Daily Gain

Sera from 354 Yorkshire and 97 Lacombe pigs on official Canadian Record of Performance test were analyzed by starch gel electrophoresis to determine their hemopexin, transferrin, prealbumin and albumin1 types. Estimates of the possible effects of different serum protein types on daily gain on test, age at slaughter adjusted to 70.37 kg carcass weight and seven other carcass characteristics were obtained by least squares methods. Significant associations were observed between each of the four serum protein loci and at least one of the nine productive traits in the Yorkshire breed. In the Lacombe breed only the prealbumin locus, which was associated with total backfat thickness, had any effect on any one of the nine productive traits. Since the results for the two breeds were not in agreement with each other and the magnitude of the difference between extremes was small, it was concluded that a correlated response in swine performance could not be expected from selection on the basis of serum protein polymorphism.

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.

scholarly journals Electron-microscopic and chemical studies of oligomers in horse ferritin

1968 ◽  
Vol 110 (2) ◽  
pp. 265-280 ◽  
Author(s):  
M. A. Williams ◽  
Pauline M. Harrison
Keyword(s):  
Electron Microscopy ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Starch Gel Electrophoresis ◽  
Quantitative Electron Microscopy ◽  
Electron Microscopic ◽  
Ph Values ◽  
Chemical Studies ◽  
Starch Gel ◽  
Intermolecular Distances

Horse ferritin was fractionated both by starch-gel electrophoresis and by gel filtration on Sephadex G-200. Monomer fractions contained up to 98% of monomer and oligomer fractions up to 76% of oligomers as determined by quantitative electron microscopy. Percentages obtained from electron micrographs correlated well with analytical starch-gel electrophoretograms and ultracentrifuge patterns. Amino acid analyses of monomer- and oligomer-enriched fractions showed no significant differences. Ferritin oligomers did not apparently dissociate on dilution for electron microscopy or on storage. Apoferritin dimers were stable in 0·01m-phosphate buffer at dilutions down to 0·19mg./ml. as shown by ultracentrifugation. Chemical studies indicated that the intermolecular bonds in oligomers are resistant to a variety of reagents and conditions, including those that would be expected to attack disulphide, peptide and ester linkages respectively. Partial disaggregation was achieved at high pH values and in 67% (v/v) acetic acid. Centre-to-centre intermolecular distances in dimers were found to be about 100å. Three main types of trimer configuration were found and a variety of tetramers and pentamers. These configurations are described and discussed.

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