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: 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.

scholarly journals Purification and characterization of hemocyte phenoloxidases in Chilo suppressalis walker (Lepidoptera: Crambidae)

2015 ◽  
Vol 67 (4) ◽  
pp. 1119-1125 ◽  
Author(s):  
Seyyedeh Mirhaghparast ◽  
Arash Zibaee ◽  
Hassan Hoda ◽  
Mahmoud Fazeli-Dinan
Keyword(s):  
Kinetic Parameters ◽  
Column Chromatography ◽  
Deae Cellulose ◽  
Chilo Suppressalis ◽  
Purification And Characterization ◽  
Molecular Weights ◽  
Specific Activities ◽  
Fast Flow ◽  
Statistical Effects

In the current study, two phenoloxidases (POs) from the larvae of Chilo suppressalis Walker were extracted and purified by column chromatography using Sepharyl G-100 and DEAE-Cellulose fast flow column. Two proteins possessing PO activity, named as POI and POII, were extracted by purification, 5.08- and 5.62-fold, respectively, with 8.94% and 7.31% recoveries, respectively. Also, the specific activities of POI and POII were 0.478 and 0.529 U/mg protein, respectively. Finally, the molecular weights of POI and POII were calculated as 94.6 and 95.7 kDa, respectively. Kinetic parameters of the purified phenoloxidases by Lineweaver-Burk analysis were Vmax of 2.27 and 1.11 U/mg protein and Km of 15.51 and 17.31 mM for POI and POII, respectively. Mg2+ and Cu2+ significantly increased the PO activities. Ca2+ decreased the activity of POI and showed no statistical effects on POII activity. EDTA and DTC significantly inhibited the activities of the purified enzymes, while triethylenetetramine hexaacetic acid (TTHA) and RGTA showed no significant effects on enzymatic activities.

PROLACTIN-LIKE ACTIVITY IN THE PITUITARY GLAND OF THE FROG

1966 ◽  
Vol 34 (2) ◽  
pp. 247-255 ◽  
Author(s):  
A. CHADWICK
Keyword(s):  
Pituitary Gland ◽  
Column Chromatography ◽  
Rana Temporaria ◽  
Sephadex Column ◽  
Zone Electrophoresis ◽  
Starch Gel ◽  
Distal Lobe

SUMMARY Homogenates of the distal lobe and combined intermediate and neural lobes from Rana temporaria and R. esculenta and extracts of these lobes, purified by zone electrophoresis in starch gel or by Sephadex column chromatography, were tested for crop-stimulating activity in the pigeon and lactogenic activity in the rabbit. The response of the pigeon crop to injections of homogenates resembled the reaction produced by purified sheep prolactin, but differed in intensity and in the number of fatty 'granules' present. The response in the rabbit differed considerably from the reaction to sheep prolactin. Milk was absent in spite of extensive proliferation in the gland. Injections of purified fractions of the pituitary extracts yielded similar results. It is concluded that a prolactin-like principle is present in the frog pituitary in both the distal lobe and the combined intermediate and neural lobes which can initiate a prolactin-like response in both the pigeon and the rabbit but cannot bring about the completion of a typical prolactin reaction.

Soluble esterases of human lung

1970 ◽  
Vol 48 (9) ◽  
pp. 970-978 ◽  
Author(s):  
J. Brebner ◽  
W. Kalow
Keyword(s):  
Molecular Weight ◽  
Column Chromatography ◽  
Human Lung ◽  
Human Tissues ◽  
Malic Dehydrogenase ◽  
Sephadex Column ◽  
Starch Gel

The soluble esterases of human lung compared to liver and serum have been illustrated on starch gel. A common zone of esterases appears to exist in nearly all human tissues and may be physiologically important acetylesterases. The most marked difference between lung and liver esterases exists in the ones moving farthest towards the anode at pH 7.0. Lactic and malic dehydrogenase patterns in the two tissues are also quite different. The zone 2 esterases appear to have a molecular weight of approximately 150 000 and those of zones 1 and 3 may be in the range of 50 000 as shown by G-200 Sephadex column chromatography.

Rhenium-188 labeled recombinant human plasminogen kringle5 (rhk5) and preliminary biodistribution

2011 ◽  
Vol 50 (06) ◽  
pp. 234-239 ◽  
Author(s):  
R. Guo ◽  
Y. Ma ◽  
R. Zhang ◽  
S. Liang ◽  
H. Shen ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Human Serum ◽  
Critical Role ◽  
Angiogenesis Inhibitor ◽  
Simple Method ◽  
Tumour Formation ◽  
Human Plasminogen ◽  
Specificity Of Binding ◽  
A549 Lung

Summary Aim: Angiogenesis plays a critical role in tumour formation and metastasis. Suitable radiolabeled angiogenesis inhibitor can be used for noninvasive imaging of angiogenesis and radionuclide therapy. Here we prepare rhenium-188 labeled recombinant human plasminogen kringle5 (188Re-rhk5) in a convenient manner than evaluate its properties in A549 lung adenocarcinoma. Methods: 188Rerhk5 was obtained by conjugating His group at the C end of rhk5 with fac- [188Re(H2O)3(CO)3]+. Chelating efficiency of fac-[188Re(H2O)3(CO)3]+ and radiolabeling efficiency of 188Re-rhk5 were measured by radio thin-layer chromatography (RTLC). In vitro stability of 188Re-rhk5 was determined in human serum at 37°C and analyzed by RTLC. Competition test was also performed to verify the specificity of binding. A biodistribution study was carried out in nude mice bearing A549 lung adenocarcinoma. Results: 188Rerhk5 was obtained with a radiolabel efficiency of 66.1%, the radiochemical purity (RCP) can marreach 95.2% after purification. 188Re-rhk5 showed high stability in human serum, the RCP was more than 80% even 12 h after incubation. Competition test showed a high binding specificity. Furthermore, this radio-complex was excreted mainly through kidneys and showed specific tumour uptake in mice bearing A549 tumours. Conclusion: 188Re-rhk5 was prepared by a simple method. Preliminary biodistribution results showed its potential as an agent for possible tumour imaging, therapy and encouraged further investigation.

Microdetermination of Cholesterol and Phospholipid in Cerebrospinal Fluid and Serum by Silicic Acid Column Chromatography

1962 ◽  
Vol 8 (6) ◽  
pp. 598-605 ◽  
Author(s):  
Yung S Shin ◽  
James C Lee
Keyword(s):  
Cerebrospinal Fluid ◽  
Human Serum ◽  
Total Phosphorus ◽  
Silicic Acid ◽  
Column Chromatography ◽  
Silicic Acid Column ◽  
Lipid Phosphorus ◽  
Silicic Acid Column Chromatography ◽  
Phospholipid Ratio

Abstract A method is presented for the determination of cholesterol and phospholipid, which requires 5 µl. of human serum or 1-2 ml. of cerebrospinal fluids. With this method 5-100 µg. of cholesterol and phospholipid can be separated by a modified silicic acid column after elution of the mixture with 1 ml. of chloroform and 3 ml. of methanol. Recovery for 24.6 µg. of cholesterol and 30.5 µg. of phospholipid was 98.4 and 96.7%, respectively. Standard deviations of ± 1.73 and ± 1.24 have been obtained for the reproducibility of cholesterol and phospholipid determinations after chromatography. The method has been applied for the estimation of the cholesterol/phospholipid ratio and of lipid phosphorus in total phosphorus of human cerebrospinal fluids.

scholarly journals Enzymic degradation of keratosulphates

1970 ◽  
Vol 119 (1) ◽  
pp. 39-47 ◽  
Author(s):  
Michiko Nishida-Fukuda ◽  
Fujio Egami
Keyword(s):  
Column Chromatography ◽  
Liver Extract ◽  
Concerted Action ◽  
Sephadex Column ◽  
Marine Gastropod ◽  
Multienzyme System ◽  
Shark Cartilage

1. A multienzyme system capable of degrading keratosulphates to yield galactose, N-acetylglucosamine and sulphate was found in the liver extract of a marine gastropod, Charonia lampas. 2. During the degradation, neither oligosaccharides nor sulphated sugars were produced. 3. It is suggested that the degradation could be attributed to the concerted action of β-galactosidase, β-N-acetylglucosaminidase and a sulphatase (sulphohydrolase), tentatively designated keratosulphatase. 4. Two forms of keratosulphatase (I and II) were separated by DEAE-Sephadex column chromatography. Both forms could release all the sulphate from keratosulphates and neither appeared to be identical with glycosulphatase or chondrosulphatase, both of which are also present in Charonia lampas. 5. β-Galactosidase and β-N-acetylglucosaminidase could degrade keratopolysulphate to a greater extent in the presence of keratosulphatase than in its absence. 6. It is suggested that keratosulphate was first desulphated by the action of keratosulphatase, and the desulphated polymer was then degraded to galactose and N-acetylglucosamine by the action of β-galactosidase and β-N-acetylglucosaminidase. 7. β-Galactosidase alone released a small amount of galactose from shark cartilage keratopolysulphate, but β-N-acetylglucosaminidase alone did not release N-acetylglucosamine. This indicates that unsulphated galactose residues occupy all the non-reducing terminal positions in keratopolysulphate chains.

Ultrasonic Effects on lsoenzymes

1966 ◽  
Vol 12 (4) ◽  
pp. 181-186 ◽  
Author(s):  
Clyde A Dubbs
Keyword(s):  
Human Serum ◽  
Ultrasonic Treatment ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Serum Cholinesterase ◽  
Starch Gel Electrophoresis ◽  
Selective Activation ◽  
Intracellular Enzymes ◽  
Starch Gel

Abstract Several significant effects of ultrasonic treatment on human serum cholinesterase and aminopeptidase isoenzymes and on other serum proteins have been found by starch gel electrophoresis. The selective activation of one cholinesterase isoenzyme is especially striking. These effects must be considered when ultrasonic treatment is used for the extraction of intracellular enzymes. When the effects are appreciated, ultrasonics should provide a valuable tool for isoenzyme research.

Sign in / Sign up

Export Citation Format

Share Document