An inducible iron-containing superoxide dismutase in Rhizobium japonicum

1981 ◽  
Vol 27 (11) ◽  
pp. 1202-1208 ◽  
Author(s):  
Mark D. Stowers ◽  
Gerald H. Elkan
Keyword(s):  
Superoxide Dismutase ◽  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Metal Content ◽  
Methyl Viologen ◽  
Polyacrylamide Gel Electrophoresis ◽  
Rhizobium Japonicum ◽  
Growth Media ◽  
Sod Activity ◽  
Rhizobium Spp

Crude extracts of Rhizobium japonicum contained a single superoxide dismutase (SOD) as revealed by polyacrylamide gel electrophoresis. Rhizobium japonicum SOD was determined to be iron-containing by criteria of sensitivity to 5 mM H2O2 and 1 mM NaN3. Growth media strongly influenced SOD activity. The addition of methyl viologen to cultures of R. japonicum 61A76NS, growing in complex medium, caused induction of the enzyme. Yeast extract was essential for induction of SOD. Puromycin blocked the induction of the enzyme. Rhizobium spp. were surveyed for SOD activity, electrophoretic mobility on polyacrylamide gels, and metal content.

scholarly journals Studies on Platelet Glycoproteins by Surface Labeling and SDS Polyacrylamide Gel Electrophoresis

1977 ◽  
Author(s):  
I. Hagen ◽  
N.O. Solum ◽  
M. Peterka
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Conformational State ◽  
Polyacrylamide Gel ◽  
The Other ◽  
Metabolic Inhibitors ◽  
Conventional System ◽  
Platelet Surface ◽  
Release Reaction

Platelet surface (glyco)proteins, and alterations in these in connection with the thrombin-induced release reaction has been studied. Platelets were labeled by lactoperoxidase-catalyzed iodination, and examined by SDS gel electrophoresis in two different gel systems, one conventional(J. Biol. Chem.1969 244 4406), and the other containing urea and EDTA in the gels. In the conventional system the bulk of radioactivity coincided with a PAS band (GP III) of MW about 100, 000. In the other system, the main radioactive peak appeared in the GP II area (MW 120,000), and a shift in the PAS stain intensity from GP III to GP II was seen. Thrombasthenic platelets showed only traces of the GP II band in both systems. The bulk of radioactivity was associated with the surface glycopolypeptide GPS, which is present, but not labeled in normal platelets. In thrombin-released platelets, GPS in its unreduced state has an altered electrophoretic mobility compared to control platelets and platelets which have been incubated with metabolic inhibitors to prevent secretion. The findings indicate that the GP III band consists of two different polypeptides, one of which appears in the GP II area in gels containing urea and EDTA. Further, that thrombasthenic platelet membrane exists in a conformational state different from that of normal platelets. And finally, GPS is in some way involved in, or influenced by, the thrombin-induced release reaction.

scholarly journals SECRETION OF PLASMINOGEN ACTIVATOR BY STIMULATED MACROPHAGES

1974 ◽  
Vol 139 (4) ◽  
pp. 834-850 ◽  
Author(s):  
Jay C. Unkeless ◽  
Saimon Gordon ◽  
E. Reich
Keyword(s):  
Amino Acids ◽  
Catalytic Activity ◽  
Plasminogen Activator ◽  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Plasminogen Activators ◽  
Peritoneal Macrophages ◽  
Chemical Labeling ◽  
Specific Catalytic Activity

Cultured thioglycollate-stimulated peritoneal macrophages synthesize, accumulate, and continuously release high levels of plasminogen activators for at least 4 days whereas cultures of unstimulated macrophages do not; the higher specific catalytic activity of released vs. cell-associated enzyme suggests that the plasminogen activators are actively secreted. The major macrophage plasminogen activator is a serine protease of mol wt 48,000, and thus resembles the comparable enzyme released by virally transformed fibroblasts. Macrophages release a second plasminogen activator of mol wt 28,000 that is also a serine enzyme. The secretion products released by stimulated and unstimulated macrophages have been compared by SDS-polyacrylamide gel electrophoresis after chemical labeling with 3H-DFP or biosynthetic labeling with 14C-amino acids. These procedures show that some proteins are formed in both cultures, whereas others are uniquely secreted by each type of macrophage. The serine enzymes released by the two kinds of macrophages differ in specificity and electrophoretic mobility.

scholarly journals Superoxide dismutase, catalase, and peroxidase in ammonium-grown and nitrogen-fixing Azospirillum brasilense

1984 ◽  
Vol 30 (10) ◽  
pp. 1222-1228 ◽  
Author(s):  
Richard W. Clara ◽  
Roger Knowles
Keyword(s):  
Superoxide Dismutase ◽  
Electron Acceptor ◽  
Azospirillum Brasilense ◽  
Polyacrylamide Gel Electrophoresis ◽  
Growth Conditions ◽  
Sod Activity ◽  
Batch Cultures ◽  
Physiological Conditions ◽  
Azospirillum Brasilense Sp7 ◽  
In Cells

Superoxide dismutase (SOD), catalase (CAT), and peroxidase (PER) activities were studied in ammonium-grown and N2-fixing batch cultures of Azospirillum brasilense Sp7. PER activity, as measured using o-dianisidine or 3,3′-diaminobenzidine as the H donor, was not significant in most growth conditions. SOD activity increased in response to higher O2 concentrations but was also present in cells grown anaerobically with nitrate [Formula: see text] or nitrous oxide (N2O) as electron acceptor. CAT activity increased at lower O2 concentrations and was highest in cells grown anaerobically with [Formula: see text] as electron acceptor. Polyacrylamide gel electrophoresis of cell-free extracts revealed only one band of SOD activity under each of the physiological conditions employed, compared with three for aerobically grown Escherichia coli K12. This band proved to be iron-containing SOD (FeSOD) on the basis of inhibitor sensitivity.

Comparison of subunits of cardiac, brain, and kidney Na+-K+-ATPase

1985 ◽  
Vol 248 (3) ◽  
pp. C247-C251 ◽  
Author(s):  
A. McDonough ◽  
C. Schmitt
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Cardiac Tissue ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Beta Subunit ◽  
Binding Affinities ◽  
Ouabain Binding ◽  
Heart Membranes ◽  
Atpase Subunits

Na+-K+-ATPase is in low abundance in cardiac tissue. Therefore, we utilized antibodies to detect the cardiac Na+-K+-ATPase subunits and to compare their characteristics with those of kidney and brain Na+-K+-ATPase subunits. By using crude preparations of heart membranes as well as purified sarcolemmal membranes from guinea pig hearts, we resolved peptides by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, blotted them onto diazotized paper, and detected Na+-K+-ATPase subunits with antibodies generated against highly purified kidney Na+-K+-ATPase holoenzyme. We tested the hypothesis that the two families of ouabain-binding affinities described in heart are due to two forms of alpha-subunit, analogous to the two forms with different affinities for ouabain described in brain. Although the antibodies did detect two forms of catalytic subunit in brain (alpha and alpha +), only one form of alpha was detected in the heart membranes, with the same electrophoretic mobility as kidney alpha. Cardiac beta-subunit could also be detected with the antikidney antibodies. It had a similar electrophoretic mobility to that described for kidney beta, whereas brain beta had a higher mobility.

Changes of Antioxidant Enzyme and Phenylalanine Ammonia-Lyase Activities during Chimonanthus praecox Seed Maturation

2008 ◽  
Vol 63 (7-8) ◽  
pp. 569-573 ◽  
Author(s):  
Xiaojia He ◽  
Shun Gao
Keyword(s):  
Superoxide Dismutase ◽  
Protein Content ◽  
Antioxidant Enzyme ◽  
Gel Electrophoresis ◽  
Growth And Development ◽  
Phenylalanine Ammonia Lyase ◽  
Polyacrylamide Gel Electrophoresis ◽  
Seed Maturation ◽  
Pal Activity ◽  
Chimonanthus Praecox

Changes in peroxidase (POD), superoxide dismutase (SOD), catalase (CAT) and phenylalanine ammonia-lyase (PAL) activities were studied during Chimonanthus praecox seed maturation. According to our findings the protein content increased steadily from 8 to 12 weeks after flowering, and thereafter decreased significantly. Similarly, SOD and POD activities increased gradually up to 12 weeks after flowering and then declined. PAL activity declined gradually during seed maturation. CAT activity, however, showed no changes during seed maturation. By means of polyacrylamide gel electrophoresis (PAGE), SOD and POD isoenzymes were observed during seed maturation. The staining intensities of SOD and POD isoenzymes correlated well with SOD and POD activities as obtained by an assay in solution. These findings suggest that POD, SOD and PAL may be involved in the growth and development during Chimonanthus praecox seed maturation.

Molecular weight determination and partial characterization of Klebsiella pneumoniae hemolysins

1986 ◽  
Vol 32 (11) ◽  
pp. 884-888 ◽  
Author(s):  
Lucila Isabel Barberis ◽  
Alberto Jorge Eraso ◽  
Maria Cristina Pàjaro ◽  
Inès Albesa
Keyword(s):  
Klebsiella Pneumoniae ◽  
Gel Electrophoresis ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Growth Media ◽  
Molecular Weight Determination ◽  
Periodic Acid Schiff

Two thiol-activated Klebsiella pneumoniae hemolysins were purified from growth media by means of salt precipitation, gel filtration, ion-exchange chromatography, and polyacrylamide gel electrophoresis. The hemolysins peaks coincided with the protein and glycoprotein peaks as determined by chromatography and electrophoresis, The molecular weights, estimated by gel filtration, were 8400 and 19 000; by sodium dodecyl sulphate–polyacrylamide gel electrophoresis, the values were calculated as 15 500 and 27 000. The electrophoretic bands were best detected by the periodic acid–Schiff method. Reduction of the disulfide linkages did not cause the originally larger molecule to break into 8400 and 19 000 hemolysins. However, trypsin treatment cleaved the 19 000 hemolysin into an active moiety, with an electrophoretic migration similar to the 8400 hemolysin. A naturally occurring proteolytic activity was investigated using pepstatin and antipain. When the trypsin inhibitor was added to the system, the hemolytic activity was detected only in the 19 000 hemolysin and the smaller hemolysin was absent.

scholarly journals Biology of amazonian mosquitoes. III. Esterase isozymes in Anopheles darlingi.

1985 ◽  
Vol 15 (1-2) ◽  
pp. 167-178 ◽  
Author(s):  
Joselita M.M. dos Santos ◽  
Eucleia P.B. Contel ◽  
Warwick E. Kerr
Keyword(s):  
Electrophoretic Mobility ◽  
Gel Electrophoresis ◽  
Developmental Stages ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Anopheles Darlingi ◽  
Genetic Studies ◽  
Esterase Isozymes ◽  
Phenotypic Distribution ◽  
Polymorphic System

Six esterase isozymes were studied during the development of Anopheles darlingi by using polyacrylamide gel electrophoresis and two different substrates, a-naphthylcelate and a-naphthylpropionate. Esterases 5 and 6'were detected in all developmental stages esterases 1 and 2 were more intensively stained if larvae, while esterases 3 and 4 were better visualized in pupae and adults. Strong differences in intensity of some of the isozymes were observed during the pupal stage.Four out of the six isozymes showed variation in the electrophoretic mobility. Esterase-2 was choosed for genetic studies, because was the best stained isozyme in the gels. Two codominant alleles {Est2*S and Est2*F) code for this polymorphic system, with the Est*S frequency equal to 0.521. Phenotypic distribution is in agreement with hardy-Weinberg expectations.

Purification Of 1- And 0-Gla Prothrombins And Their Differentiation With Normal And Other Dicoumarol-Induced Prothrombins

1981 ◽  
Author(s):  
O P Malhotra
Keyword(s):  
Sodium Dodecyl Sulfate ◽  
Electrophoretic Mobility ◽  
Vitamin K ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
The Other ◽  
Multiple Forms ◽  
Isoelectric Precipitation ◽  
Antigenic Activity

In normal prothrombin, 10-glutamyl residues present in the amino portion of the molecule are carboxylated to form γ-carboxyglutamic acids (gla). Dicoumarol, an antagonist of vitamin K, induces the production of (partially) acarboxylated atypical prothrombins. In addition to our atypical varieties, viz. 7−,5− and 2-gla prothrombins, we have isolated two more atypical molecules, one containing 1.0±0.1 gla (1-gla prothrombin) and the other approximately 0.25 gla (0-gla prothrombin). These two variants adsorbed onto alumina Cγ-gel (Bio-Rad), similar to 2-gla protein, but were derived from 40 to 50% (NH4)2SO4 saturation. The purified materials, obtained after isoelectric precipitation followed by preparatory polyacrylamide-gel electrophoresis and heparin agarose chromatography, showed a single component by analytical disc-gel electrophoresis both in the presence or absence of sodium dodecyl sulfate (SDS) and contained antigenic activity comparable to that of normal prothrombins.The pI’s of the two variants by column electrofocusing were each 4.835±0.015. Similarly, the two proteins did not reveal any difference in electrophoretic mobility; however, their prothrombin fragments 1 (F1, residues 1-156) did—0-gla F1 moved slower than 1-gla F1. Employing anti (normal) prothrombin sera with Ca2+ , the two, 0− and 1-gla, F1’s produced vaguely visible immunoprecipitates which were definitely lighter than all the other F1’s including 2-gla. These results confirm that not only are multiple forms of atypical prothrombin induced by dicoumarol but also that gla does affect the immunochemical properties of the gla-containing fragment.

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