scholarly journals Isolation and characterization of glyoxylate dehydrogenase from the fungus Sclerotium rolfsii

1984 ◽  
Vol 218 (1) ◽  
pp. 113-118 ◽  
Author(s):  
A J Balmforth ◽  
A Thomson
Keyword(s):  
Affinity Chromatography ◽  
Active Site ◽  
Sclerotium Rolfsii ◽  
Thiol Group ◽  
Isolation And Characterization ◽  
Crude Extracts ◽  
Nad Oxidoreductase ◽  
Inhibition Studies ◽  
Dye Affinity Chromatography

Glyoxylate dehydrogenase (glyoxylate: NAD+ oxidoreductase) was purified 600-fold in three steps from crude extracts of the fungus Sclerotium rolfsii (Corticium rolfsii Curzi). Two of the purification steps involved dye-affinity chromatography. The enzyme is a tetramer of Mr 250 000, with identical subunits of Mr 57 000. Inhibition studies suggest that there is one essential thiol group per active site.

scholarly journals The isolation and characterization of 3-phosphoglycerate dehydrogenase from peas

1968 ◽  
Vol 109 (5) ◽  
pp. 743-748 ◽  
Author(s):  
J. C. Slaughter ◽  
D. D. Davies
Keyword(s):  
Thiol Group ◽  
Isolation And Characterization ◽  
Crude Extracts ◽  
Michaelis Constants ◽  
Phosphoglycerate Dehydrogenase

1. 3-Phosphoglycerate dehydrogenase was purified 400-fold from crude extracts of etiolated pea epicotyls. 2. Michaelis constants were determined for all four substrates. 3. Loss of sensitivity to inhibition by l-serine occurs on purification. 4. The purified enzyme is inhibited by thiol-group reagents and, with N-ethyl-maleimide, protection is afforded by 3-phosphoglycerate though not by NAD+.

Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

1974 ◽  
Vol 31 (01) ◽  
pp. 072-085 ◽  
Author(s):  
M Kopitar ◽  
M Stegnar ◽  
B Accetto ◽  
D Lebez
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Gel Electrophoresis ◽  
Gel Filtration ◽  
Ph Optimum ◽  
Isolation And Characterization ◽  
Ph Range ◽  
Inhibition Studies ◽  
Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

scholarly journals Isolation and characterization of a low-molecular-weight substance activating head and bud formation in hydra

Development ◽  
1973 ◽  
Vol 29 (1) ◽  
pp. 27-38
Author(s):  
H. Chica Schaller
Keyword(s):  
Molecular Weight ◽  
Low Molecular Weight ◽  
Bud Formation ◽  
Isolation And Characterization ◽  
Crude Extracts ◽  
Head Formation ◽  
Weight Substance

From crude extracts of hydra, a substance activating head formation was isolated and enriched at least 100000-fold. The molecular weight was determined to be approximately 900. Sensitivity against proteases suggests that it is a peptide. The substance acts at a concentration equivalent to the extract of 1 hydra per ml or at a concentration of less than 10 10M. In its highly purified form the substance activates head and bud formation.

scholarly journals Isolation and characterization of a digoxin-like immunoreactive substance from human urine by affinity chromatography

1997 ◽  
Vol 43 (8) ◽  
pp. 1416-1420 ◽  
Author(s):  
Claudio De Angelis ◽  
Massimiliano Riscazzi ◽  
Riccardo Salvini ◽  
Alfonso Piccoli ◽  
Claudio Ferri ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Human Urine ◽  
Biological Fluids ◽  
Active Material ◽  
Reversed Phase ◽  
Cross Reactivity ◽  
Linear Gradient ◽  
Isolation And Characterization ◽  
Dog Kidney

Abstract A series of observations has suggested that one or more digoxin-like immunoreactive substances (DLIS) in biological fluids is able to cross-react with the antidigoxin antibody. Whether this substance is the endogenous inhibitor of Na+/K+ ATPase has not been well established. The aim of this study was to identify and characterize DLIS from human urine. Treated urine from healthy men was run on an affinity chromatography column at a flow rate of 1 mL/min in which the ligand was an antibody (antiserum) to digoxin. Eluates from affinity chromatography were applied onto analytical reversed-phase HPLC. The active material was eluted with a linear gradient of acetonitrile (from 350 to 650 mL/L) and water. A second step in HPLC was carried out isocratically with 280 mL/L acetonitrile in water. We found a single peak showing cross-reactivity with antidigoxin antibody as measured by RIA. It showed the same retention time as that of a digoxin calibrator. This highly purified substance is able to displace [3H]ouabain from dog kidney-derived Na+/K+ ATPase, to inhibit Na+/K+ ATPase activity as measured by the86Rb+ uptake in red blood cells and by coupled enzyme assay. Our results are consistent with the hypothesis that DLIS, as isolated by this particular digoxin antibody, is a single substance and an inhibitor of Na+/K+ ATPase.

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