scholarly journals Some properties of active and latent catechol oxidase of mushroom

2013 ◽  
Vol 52 (1-2) ◽  
pp. 139-147 ◽  
Author(s):  
Janusz Czapski
Keyword(s):  
Kinetic Data ◽  
Sodium Dodecyl ◽  
Electrophoretic Analysis ◽  
Inhibitory Effect ◽  
Catechol Oxidase ◽  
Latent Form ◽  
Salicyl Hydroxamic Acid ◽  
Slow Moving ◽  
Salicyl Hydroxamic ◽  
Catalytic Power

Latent form of mushroom catechol oxidase was activated by O,1% sodium dodecyl sulfate (SDS). Catalytic power of the latent form, calculated from the kinetic parameters was 1,8 times higher than that of active one. Salicyl hydroxamic acid (SHAM) appeared as a powerful inhibitor for both active and latent forms of catechol oxidase. However, in the range of 150-250 μM SHAM the inhibitory effect for active catechol oxidase was significantly higher than that for the latent one. Non-competitive and irreversible characteristics of inhibition of latent and active catechol oxidase was calculated from kinetic data. Electrophoretic analysis followed by scanning of the gels was used. The spots' absorbance was determined from a computer image of the isoenzyme band patterns. It allowed us to estimate gels quantitatively. Presence of one additional clearly defined slow moving isoform of SDS-activated catechol oxidase, differed in the respect of 3 bands for the active and 4 bands for the total.

scholarly journals Some characteristics of active and latent monophenolase of mushroom polyphenol oxidase

2013 ◽  
Vol 51 (1-2) ◽  
pp. 33-41 ◽  
Author(s):  
Janusz Czapski
Keyword(s):  
Polyphenol Oxidase ◽  
Quantitative Estimation ◽  
Active Form ◽  
Sodium Dodecyl ◽  
Maximum Velocity ◽  
Electrophoretic Analysis ◽  
Qualitative And Quantitative ◽  
Monophenolase Activity ◽  
Lag Period ◽  
Catalytic Power

Latent form of monophenolase of mushroom polyphenol oxidase (PPO) was activated by 0,1% sodium dodecyl sulfate (SDS). The addition of increasing concentrations of 4-methylcatechol di minished lag period of active and total monophenolase activity, measured using p-cresol with L-proline as a substrate. Changes of lag period were described by equation of one phase exponential decay when concentration of substrate varied from l to 10 mM. Affinity ( 1/K<sub>m</sub>) toward substrate of latent monophenolase was over two times higher than that of the active form, while the maximum velocity (V<sub>max</sub>) was two times lower. The catalytic power (V<sub>max</sub>/K<sub>m</sub>) of both forms of monophenolase were almost equal. Electrophoretic analysis followed by scanning technique of the gels was used. Absorbancy of spots, determined from computer image of isoenzyme bands pattern allowed for qualitative and quantitative estimation of electrophoregrams. Presence of one additional clearly defined slow migrating isoenzyme for SDS activated monophenolase differed in this respect active (2 bands) and total (3 bands) forms of monophenolase.

Flotation Behavior and Mechanism of Anglesite with Salicyl Hydroxamic Acid as Collector

JOM ◽  
2018 ◽  
Vol 70 (12) ◽  
pp. 2813-2818 ◽  
Author(s):  
Wei Yao ◽  
Maolin Li ◽  
Rui Cui ◽  
Xingke Jiang ◽  
Hongqiang Jiang ◽  
...  
Keyword(s):  
Hydroxamic Acid ◽  
Salicyl Hydroxamic Acid ◽  
Salicyl Hydroxamic ◽  
Flotation Behavior

Preliminary Treatment of Urinary Proteins Improves Electrophoretic Analysis and Immunodetection

1992 ◽  
Vol 38 (6) ◽  
pp. 860-863 ◽  
Author(s):  
J M Verdier ◽  
B Dussol ◽  
P Dupuy ◽  
Y Berland ◽  
J C Dagorn
Keyword(s):  
Protein Pattern ◽  
Protein A ◽  
Sodium Dodecyl ◽  
Complex Mixture ◽  
Electrophoretic Analysis ◽  
Renal Physiology ◽  
Preliminary Treatment ◽  
Urinary Proteins ◽  
Simple Method ◽  
Electrophoretic Migration

Abstract Analysis of urinary protein composition is an important tool in studies on renal physiology and physiopathology. Urine is, however, a complex mixture containing, besides protein, a variety of compounds such as salts, peptides, oligosaccharides, and glycosaminoglycans. Some of these compounds interfere with the electrophoretic migration of protein in sodium dodecyl sulfate-polyacrylamide gels and prevent correct analysis of the protein pattern. We describe a simple method for extracting urinary proteins that considerably improves their electrophoretic migration and subsequent immunodetection. This treatment involves ammonium sulfate fractionations (for precipitating proteins), EDTA (for inhibiting protein aggregation), and HCl hydrolysis (for removing glycosylaminoglycans). Recovery during extraction was found to be almost quantitative for total protein and three representative proteins: albumin, alpha 1-glycoprotein acid, and beta 2-microglobulin.

Two-dimensional electrophoretic analysis of cytosols from human breast tumors: optimal migration conditions.

1984 ◽  
Vol 30 (12) ◽  
pp. 1947-1949 ◽  
Author(s):  
P Motté ◽  
J M Bidart ◽  
J C Delarue ◽  
E Comoy ◽  
P Moingeon ◽  
...  
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Breast Tumors ◽  
Electrophoretic Analysis ◽  
Ph Gradient ◽  
Two Dimensional ◽  
Human Breast ◽  
Cytosolic Proteins ◽  
Optimal Migration ◽  
Dodecyl Sulfate

Abstract In an examination of cytosols from human breast tumors, we performed two-dimensional electrophoretic analysis. Several migration conditions were tried in a search for a homogeneous repartition of cytosolic proteins. The most workable gels were obtained with a 4.40 to 8.05 pH gradient in the first dimension and a homogeneous 125 g/L acrylamide/sodium dodecyl sulfate gel, 1 mm in thickness, in the second dimension. The practicability of this method is discussed.

Inhibition of cardiac sarcolemmal Na+/H+ antiporter by opioids

1992 ◽  
Vol 70 (7) ◽  
pp. 1048-1056 ◽  
Author(s):  
S. M. Periyasamy
Keyword(s):  
Opioid Receptors ◽  
Kinetic Data ◽  
Direct Interaction ◽  
Inhibitory Effect ◽  
Transport Systems ◽  
Opioid Agonist ◽  
Inhibitory Potency ◽  
Cardiac Sarcolemma ◽  
Analog Ic ◽  
Sarcolemmal Vesicles

In our routine screening of chemicals that would inhibit cardiac sarcolemmal Na+/H+ antiporter, we discovered that some of the opioids produced inhibition of cardiac sarcolemmal Na+/H+ antiporter in micromolar concentrations. Using U-50,488H, a selective κ-opioid agonist, we characterized the nature of interaction between opioids and the Na+/H+ antiporter. The inhibitory effect of U-50,488H on Na+/H+ antiporter was immediate and reversible, and was not mediated through the interaction with the opioid receptors but due to the direct interaction of U-50.488H with the Na+/H+ antiporter. The kinetic data show that in the presence of U-50,488H the Km for Na+ was increased from 2.5 ± 0.2 to 5.0 ± 0.3 mM, while the Vmax (52.0 ± 5.0 nmol∙mg−1∙min−1) remained the same. These results suggest that U-50,488H and Na+ compete for the same site on the antiporter. When testing the effect of U-50,488H on other transport systems of cardiac sarcolemma, we found that U-50,488H also inhibited Na+/Ca2+ antiporter and Na+/K+ pump but at much higher concentrations suggesting that U-50,488H shows some degree of selectivity for cardiac sarcolemmal Na+/H+ antiporter. When we compared the inhibitory potency of U-50,488H with amiloride and its analog, namely 5-(N,N-hexamethylene)amiloride, we found that U-50,488H (IC50 = 100 ± 15 μM) was threefold more potent than amiloride (IC50 = 300 ± 20 μM) but it was threefold less potent than the amiloride analog (IC50 = 30 ± 10 μM) in inhibiting cardiac sarcolemmal Na+/H+ antiporter. These results show that although U-50,488H is more potent than amiloride, the inhibitory characteristics of U-50,488H on cardiac sarcolemmal Na+/H+ antiporter are similar to amiloride.Key words: Na+/H+ antiporter, Na+ uptake, cardiac sarcolemmal vesicles, opioid agonists, U-50,488H.

scholarly journals Simple efficient methods for the isolation of malate dehydrogenase from thermophilic and mesophilic bacteria

1979 ◽  
Vol 177 (2) ◽  
pp. 441-448 ◽  
Author(s):  
I P Wright ◽  
T K Sundaram
Keyword(s):  
Affinity Chromatography ◽  
Malate Dehydrogenase ◽  
Sodium Dodecyl Sulphate ◽  
Sodium Dodecyl ◽  
Electrophoretic Analysis ◽  
Polyacrylamide Gel ◽  
Moderately Thermophilic ◽  
Extreme Thermophiles ◽  
Moderate Thermophiles ◽  
Number Of Bacteria

Malate dehydrogenase from a number of bacteria drawn from several genera and representing the mesophilic, moderately thermophilic and extremely thermophilic classes was isolated by procedures which involve only a small number of steps (in most cases only two), of which the key one is affinity chromatography on 5′-AMP–Sepharose and/or on NAD+–hexane–agarose. Electrophoretic analysis of the native enzymes in polyacrylamide gel and of the denaturated enzymes in sodium dodecyl sulphate/polyacrylamide gel revealed no significant protein impurity in the purified preparations. The yields ranged from about 40% to over 80%. The malate dehydrogenases from the extreme thermophiles and from some of the moderate thermophiles are appreciably less efficient catalytically than their mesophilic homologues.

Comparative modeling of the latent form of a plant catechol oxidase using a molluskan hemocyanin structure

2002 ◽  
Vol 89 (1-2) ◽  
pp. 155-158 ◽  
Author(s):  
Carsten Gerdemann ◽  
Christoph Eicken ◽  
Hans-Joachim Galla ◽  
Bernt Krebs
Keyword(s):  
Comparative Modeling ◽  
Catechol Oxidase ◽  
Latent Form

scholarly journals The lanthanide-enhanced affinity chromatography of clostridial collagenase

1985 ◽  
Vol 225 (2) ◽  
pp. 553-556 ◽  
Author(s):  
C H Evans
Keyword(s):  
Periodic Acid ◽  
Sodium Dodecyl ◽  
Minor Component ◽  
Electrophoretic Analysis ◽  
Lanthanide Ions ◽  
Affinity Column ◽  
Major Protein ◽  
Periodic Acid Schiff ◽  
Highly Active ◽  
A Minor

Clostridiopeptidase A (EC 3.4.24.3) did not bind to a collagen affinity column in the absence of Ca2+, but did so in the presence of lanthanide ions (Ln3+). The sequestered enzyme could be eluted with EGTA. For the four Ln3+ ions tested, the order of efficiency in promoting enzyme binding, Sm3+ greater than Lu3+ greater than Er3+ much greater than La3+, reflected their relative abilities to inhibit clostridiopeptidase A. By using Sm3+ as an adjunct, it proved possible to separate a highly active preparation of collagenase from crude clostridial collagenase. Sodium dodecyl sulphate/polyacrylamide-gel-electrophoretic analysis of the preparation revealed a major protein of Mr 95000 and a minor component of Mr 82000. As both were stained by periodic acid/Schiff reagent, they were probably glycoproteins.

Electrophoretic Analysis of Polypeptides of Plasma Membranes from Bovine Pituitary Gland

1974 ◽  
Vol 52 (7) ◽  
pp. 620-630
Author(s):  
André Lemay ◽  
Fernand Labrie
Keyword(s):  
Plasma Membranes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Broad Band ◽  
Sodium Dodecyl ◽  
Electrophoretic Pattern ◽  
Apparent Molecular Weight ◽  
Electrophoretic Analysis ◽  
Polyacrylamide Gel ◽  
Protein Components ◽  
Membrane Components

Purified plasma membranes from bovine hypophyseal tissue have been fractionated by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis under various conditions of pH and acrylamide concentrations. The best separation of protein components is achieved at a concentration of 7.5% acrylamide and at pH 7.1. Under these conditions, the electrophoretic pattern consistently shows 36 protein bands ranging in molecular weights from 250 000 to 15 000. Only one broad band, having an apparent molecular weight of 150 000, stains for glycoproteins by the period acid – Schiff technique. After electrophoresis on a two-dimensional polyacrylamide gel system using disc gels containing urea and Triton X-100 in the first dimension and SDS in the second dimension, approximately 45 different protein components can be identified. Less than 12% of the membrane proteins are solubilized by washing the membranes with 1 M KCl or NH4Cl. Denaturating agents like urea and lithium 3,4-diiodosalycilate solubilize 55–60% of membrane components. Adenohypophyseal plasma membranes show an eleetrophoretic pattern completely different from that obtained with membranes isolated from the intermediate or posterior pituitary lobes.

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