Solubilization, Partial Purification, and Affinity Labeling of the Membrane-Bound Isoprenylated Protein Endoprotease†

Biochemistry ◽  
1996 ◽  
Vol 35 (10) ◽  
pp. 3227-3237 ◽  
Author(s):  
Yulong Chen ◽  
Yu-ting Ma ◽  
Robert R. Rando
Keyword(s):  
Partial Purification ◽  
Affinity Labeling ◽  
Membrane Bound

scholarly journals MICROSOMAL MEMBRANE-BOUND SERINE PROTEINASE FROM RAT LIVER: PARTIAL PURIFICATION AND SPECIFICITY TOWARD ARGINYL PEPTIDE BONDS

1991 ◽  
Vol 12 (2) ◽  
pp. 113-120 ◽  
Author(s):  
MITSUAKI YANAGIDA ◽  
YOSHIAKI TAMANOUE ◽  
KAZUO SUTOH ◽  
TAKAYUKI TAKAHASHI ◽  
KENJI TAKAHASHI
Keyword(s):  
Rat Liver ◽  
Serine Proteinase ◽  
Microsomal Membrane ◽  
Partial Purification ◽  
Peptide Bonds ◽  
Membrane Bound

Partial purification and characterization of a new human membrane-bound carbonyl reductase playing a role in the deactivation of the anticancer drug oracin

Toxicology ◽  
2009 ◽  
Vol 264 (1-2) ◽  
pp. 52-60 ◽  
Author(s):  
Lucie Škarydová ◽  
Adam Skarka ◽  
Romana Novotná ◽  
Lucie Živná ◽  
Hans-Jörg Martin ◽  
...  
Keyword(s):  
Anticancer Drug ◽  
Carbonyl Reductase ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Membrane Bound

scholarly journals Interference of chlorate and chlorite with nitrate reduction in resting cells of Paracoccus denitrificans

Microbiology ◽  
2006 ◽  
Vol 152 (12) ◽  
pp. 3529-3534 ◽  
Author(s):  
Igor Kučera
Keyword(s):  
Cell Membrane ◽  
Nitrate Reductase ◽  
Nitrate Reduction ◽  
Paracoccus Denitrificans ◽  
Metabolic Activation ◽  
Partial Purification ◽  
Resting Cells ◽  
Aerobic Growth ◽  
Membrane Bound ◽  
Periplasmic Enzyme

When grown anaerobically on a succinate+nitrate (SN) medium, Paracoccus denitrificans forms the membrane-bound, cytoplasmically oriented, chlorate-reducing nitrate reductase Nar, while the periplasmic enzyme Nap is expressed during aerobic growth on butyrate+oxygen (BO) medium. Preincubation of SN cells with chlorate produced a concentration-dependent decrease in nitrate utilization, which could be ascribed to Nar inactivation. Toluenization rendered Nar less sensitive to chlorate, but more sensitive to chlorite, suggesting that the latter compound may be the true inactivator. The Nap enzyme of BO cells was inactivated by both chlorate and chlorite at concentrations that were at least two orders of magnitude lower than those shown to affect Nar. Partial purification of Nap resulted in insensitivity to chlorate and diminished sensitivity to chlorite. Azide was specific for SN cells in protecting nitrate reductase against chlorate attack, the protective effect of nitrate being more pronounced in BO cells. The results are discussed in terms of different metabolic activation of chlorine oxoanions in both types of cells, and limited permeation of chlorite across the cell membrane.

scholarly journals Partial Purification of Membrane-bound b-Type Cytochrome from Halobacterium halobium.

1979 ◽  
Vol 33b ◽  
pp. 600-601 ◽  
Author(s):  
Christina Hallberg ◽  
Herrick Baltscheffsky ◽  
Olle Karlström ◽  
Toshiaki Nishida ◽  
Curt R. Enzell ◽  
...  
Keyword(s):  
Partial Purification ◽  
Halobacterium Halobium ◽  
Membrane Bound

Separation, Partial Purification and Characterization of a Fatty Acid Hydroperoxide Cleaving Enzyme from Apple and Tomato Fruits

1982 ◽  
Vol 37 (3-4) ◽  
pp. 165-173 ◽  
Author(s):  
P. Schreier ◽  
G. Lorenz
Keyword(s):  
Fatty Acid ◽  
Inhibitory Effect ◽  
Fatty Acid Hydroperoxide ◽  
Partial Purification ◽  
Purification And Characterization ◽  
Ph Optimum ◽  
Isoelectric Points ◽  
Acid Hydroperoxide ◽  
Membrane Bound

Abstract A membrane-bound enzyme catalysing the cleavage of 13-hydroperoxy-(Z)-9,(E)-11-oc-tadecadienoic acid (13-LHPO) and 13-hydroperoxy-(Z)-9,(E)-11,(Z)-15-octadecadienoic acid (13-LnHPO) to C6-aldehydes was isolated and partially purified from apples and tomatoes. Attempts to employ Ultrogel AcA 34 and AcA 22 in a gel chromatographic purification step were partially frustrated by reaggregation phenomena. However, by using Sepharose CL-4 B an enzyme fraction (MW 200 000 Da) with lipoxygenase and fatty acid hydroperoxide cleaving activity could be separated from a high molecular-weight active eluate. By applying preparative isoelec­ tric focussing to the tomato protein we succeeded in separating the fatty acid cleaving activity from the lipoxygenase, because o f their different isoelectric points of pH 5.8 -6 .1 and pH 5.0, respectively, An 8.4-fold purification of the fatty acid cleaving activity was achieved. A pH-optimum of 5.5 and a Km-value of 2.6 × 10-5 м/1 for the 13-hydroperoxide of linoleic acid were measured. p-Chloromercuribenzoic acid (1 mм) showed significant inhibitory effect on the fatty acid hydroperoxide cleaving enzyme, but no evidence o f inhibition was found with 1 mм H2O2, KCN, DABCO and EDTA or superoxide dismutase (270 U). The maximum amount of fatty acid hydroperoxide decomposition (C6-aldehyde formation) was determined to be 59%.

Adenylyl Cyclase from the Green Alga Volvox carteri f . nagariensis: Partial Purification and Characterisation

1994 ◽  
Vol 21 (5) ◽  
pp. 613 ◽  
Author(s):  
N Nass ◽  
R Moka ◽  
L Jaenicke
Keyword(s):  
Enzyme Activity ◽  
Adenylyl Cyclase ◽  
Biochemical Properties ◽  
Partial Purification ◽  
Volvox Carteri ◽  
Affinity Labelling ◽  
Membrane Bound ◽  
Low Levels ◽  
Atp Binding Protein ◽  
Plant Enzyme

In order to understand changes in cyclic adenylate levels of Volvox carteri during the process of sexual induction, we investigated the biochemical properties of its membrane-bound adenylyl cyclase. Membrane preparations possess low levels of Mg2+ -dependent or Mn2+ -dependent adenylyl cyclase activity. This activity was solubilised and then purified 7800-fold. The enzyme detergent complex has an apparent molecular mass of 100 kDa. Purified preparations contain a major ATP-binding protein of 33 kDa as shown by affinity labelling. The Mg2+ -dependent basal enzyme activity is regulated by Ca2+, and is highest in the presence of 10-7 M Ca2+, but is inhibited by Ca2+ above 10-5 M. La3+ at 10-4M also blocks activity. Neither calmodulin nor its antagonists affect the enzyme activity, nor do the purified preparations interact with immobilised calmodulin. Further mediators of G-protein action (NaF, or GTP and its derivatives) and forskolins have no influence on the basal activity of this plant enzyme. The function of adenylyl cyclase in sexual induction of Volvox is discussed.

scholarly journals Partial purification and characterization of a peroxidase activity from human placenta

1990 ◽  
Vol 268 (3) ◽  
pp. 739-743 ◽  
Author(s):  
J L Nelson ◽  
A P Kulkarni
Keyword(s):  
Peroxidase Activity ◽  
Potential Role ◽  
Gel Filtration ◽  
Partial Purification ◽  
Human Haemoglobin ◽  
Prostaglandin Synthase ◽  
Membrane Bound ◽  
Cytochrome P 450

Peroxidases can metabolize a variety of xenobiotics to reactive intermediates capable of binding to protein or DNA. The potential role of these enzymes in fetotoxicity has not been explored. In this study, the presence of peroxidase activity was observed in human term and pre-term placenta. Human term placental peroxidase activity (HTPP) was partially purified by concanavalin A affinity chromatography from CaCl2 extracts of the particulate fraction. HTPP appears to be a membrane-bound glycoprotein. Arachidonic acid-dependent oxidation of guaiacol was not observed, suggesting that the peroxidase activity was not due to prostaglandin synthase. Moreover, HTPP preparations were devoid of catalase and spectrally dissimilar from human haemoglobin, cytochrome P-450, eosinophil peroxidase and myloperoxidase, suggesting an endogenous origin. An Mr of approx. 119,000 was determined for HTPP by gel filtration. Cathodic slab-PAGE of cetyltrialkylammonium bromide-solubilized HTPP yielded two peroxidase-staining bands.

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