Hemolytic and Elastolytic Activities Influenced by Iron in Plesiomonas shigelloides

1999 ◽  
Vol 62 (12) ◽  
pp. 1475-1477 ◽  
Author(s):  
JESÚS A. SANTOS ◽  
CÉSAR J. GONZÁLEZ ◽  
TERESA M. LÓPEZ ◽  
ANDRÉS OTERO ◽  
MARÍA-LUISA GARCÍA-LÓPEZ
Keyword(s):  
Thermal Treatment ◽  
Hemolytic Activity ◽  
Serine Proteases ◽  
Culture Media ◽  
Plesiomonas Shigelloides ◽  
Elastin Degradation ◽  
Elastolytic Activity ◽  
Degradation Activity ◽  
Phenylmethyl Sulfonyl Fluoride ◽  
Sulfonyl Fluoride

The aim of this study was to determine the presence of hemolytic and elastolytic enzymes in several strains of Plesiomonas shigelloides in relation to the availability of iron in culture media. Hemolytic activity and elastolytic activity were detected in strains of P. shigelloides and were enhanced when the strains were grown in an iron-depleted medium and lost after thermal treatment at 100°C for 10 min. Also, elastolytic activity was inactivated by phenylmethyl sulfonyl fluoride, an inhibitor of serine proteases. Hemolytic activity was detected extracellularly in cell-free supernatants, whereas elastin degradation activity was cell associated. Both activities may be related to the virulence of P. shigelloides.

scholarly journals Co-operation between plasmin and elastase in elastin degradation by intact murine macrophages

1984 ◽  
Vol 222 (3) ◽  
pp. 721-728 ◽  
Author(s):  
H A Chapman ◽  
O L Stone
Keyword(s):  
Conditioned Medium ◽  
Proteinase Inhibitors ◽  
Intact Cell ◽  
Cysteine Proteinase ◽  
Cell Activity ◽  
Live Cells ◽  
Murine Macrophages ◽  
Elastin Degradation ◽  
Elastolytic Activity ◽  
Cell Conditioned Medium

Intact, thioglycollate-stimulated murine macrophages cultured on an insoluble [3H]-elastin substratum progressively hydrolysed the elastin. Cell lysates had little activity. We compared the effect of various proteinase inhibitors on elastinolysis by either live cells or cell-free, elastase-rich conditioned medium. Only known inhibitors of macrophage elastase blocked the activity of elastase-rich cell-conditioned medium whereas inhibitors of cathepsin B also suppressed intact cell activity. Serum proteinase inhibitors blocked cell-derived soluble elastase activity but not intact cell elastolytic activity. We also observed that plasminogen added to the cell cultures markedly increased elastinolysis by live macrophages or cell-free elastase-rich medium. Purified plasmin alone had no measurable effect on native elastin. Additional experiments indicated that the plasmin enhancement was due to elastin-dependent activation of latent macrophage elastase. These results indicate that live macrophage elastinolysis is a co-operative process involving multiple proteinases, especially a cysteine proteinase(s) and elastase. Plasmin may be a physiological activator of latent macrophage elastase.

scholarly journals Extracellular Polyhydroxyalkanoate Depolymerase by Acidovorax sp. DP5

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
S. Vigneswari ◽  
T. S. Lee ◽  
Kesaven Bhubalan ◽  
A. A. Amirul
Keyword(s):  
Biochemical Characterization ◽  
Culture Media ◽  
Mineral Salt Medium ◽  
Alkaline Condition ◽  
Degradation Index ◽  
Cloning And Sequencing ◽  
Assay Condition ◽  
Carbon And Nitrogen ◽  
Water And Soil Samples ◽  
Degradation Activity

Bacteria capable of degrading polyhydroxyalkanoates (PHA) by secreting extracellular depolymerase enzymes were isolated from water and soil samples collected from various environments in Malaysia. A total of 8 potential degraders exhibited clear zones on poly(3-hydroxybutyrate) [P(3HB)] based agar, indicating the presence of extracellular PHA depolymerase. Among the isolates, DP5 exhibited the largest clearing zone with a degradation index of 6.0. The highest degradation activity of P(3HB) was also observed with depolymerase enzyme of DP5 in mineral salt medium containing P(3HB). Based on biochemical characterization and 16S rRNA cloning and sequencing, isolate DP5 was found to belong to the genus Acidovorax and subsequently named as Acidovorax sp. DP5. The highest extracellular depolymerase enzyme activity was achieved when 0.25% (w/v) of P(3HB) and 1 g/L of urea were used as carbon and nitrogen source, respectively, in the culture media. The most suitable assay condition of the depolymerase enzyme in response to pH and temperature was tested. The depolymerase produced by strain Acidovorax sp. DP5 showed high percentage of degradation with P(3HB) films in an alkaline condition with pH 9 and at a temperature of 40°C.

Cell-Associated Hemolytic Activity in Environmental Strains of Plesiomonas shigelloides Expressing Cell-Free, Iron-Influenced Extracellular Hemolysin

2007 ◽  
Vol 70 (4) ◽  
pp. 885-890 ◽  
Author(s):  
NIEVES GONZÁLEZ-RODRÍGUEZ ◽  
JESÚS A. SANTOS ◽  
ANDRÉS OTERO ◽  
MARÍA-LUISA GARCÍA-LÓPEZ
Keyword(s):  
Hemolytic Activity ◽  
Pathogenic Bacteria ◽  
Total Activity ◽  
Growth Conditions ◽  
Heme Iron ◽  
Outer Membrane Receptor ◽  
Plesiomonas Shigelloides ◽  
Testing Procedures ◽  
A Cell

Hemolysis is a means of providing pathogenic bacteria with heme iron in vivo. In a previous work, iron-influenced hemolytic activity against sheep erythrocytes was detected in cell-free supernatants, but not in the cell fraction of two environmental Plesiomonas shigelloides strains incubated without shaking. Both strains have the hugA gene, which encodes an outer membrane receptor required for heme iron utilization. The present study was undertaken to investigate the expression of a second hemolytic activity detected during aerated incubation in normal and iron-depleted tryptone soya broth (id-TSB). An agar overlay procedure and doubling dilution titrations were employed to detect the hemolytic activity against several erythrocyte species. The kinetics of growth and hemolytic activity were assayed at 35°C in aerated normal and id-TSB and salmon extract. Overlaid colonies showed a cell-associated beta-hemolytic activity within 4 h. For aerated cell-free supernatants, titers above 16 were not attained until 30 to 48 h of incubation; the best activity was noted with dog and mouse erythrocytes. After 24 h of aerated incubation, sonicated cells yielded high hemolytic activity against dog erythrocytes without activity in supernatants, but after 48 h, only 28 to 30% of the total activity remained cell associated. The hemolytic factor was released in broths during the death phase. Hemolytic activity was not detected in fish extract. This and other studies suggest that P. shigelloides may produce at least two hemolytic factors, their expression and detection being influenced by environmental growth conditions and testing procedures. The overlay assay appears to be the best routine method for detecting hemolytic activity in P. shigelloides.

scholarly journals Characterization of cathepsin S exosites that govern its elastolytic activity

2020 ◽  
Vol 477 (1) ◽  
pp. 227-242 ◽  
Author(s):  
Pierre-Marie Andrault ◽  
Preety Panwar ◽  
Dieter Brömme
Keyword(s):  
Binding Site ◽  
Structural Features ◽  
Docking Studies ◽  
Structural Examination ◽  
Structural Localization ◽  
Elastin Degradation ◽  
Basement Membrane Component ◽  
Elastolytic Activity ◽  
The Right

We have previously determined that the elastolytic activities of cathepsins (Cat) K and V require two exosites sharing the same structural localization on both enzymes. The structural features involved in the elastolytic activity of CatS have not yet been identified. We first mutated the analogous CatK and V putative exosites of CatS into the elastolytically inactive CatL counterparts. The modification of the exosite 1 did not affect the elastase activity of CatS whilst mutation of the Y118 of exosite 2 decreased the cleavage of elastin by ∼70% without affecting the degradation of other macromolecular substrates (gelatin, thyroglobulin). T06, an ectosteric inhibitor that disrupt the elastolytic activity of CatK, blocked ∼80% of the elastolytic activity of CatS without blocking the cleavage of gelatin and thyroglobulin. Docking studies showed that T06 preferentially interacts with a binding site located on the Right domain of the enzyme, outside of the active site. The structural examination of this binding site showed that the loop spanning the L174N175G176K177 residues of CatS is considerably different from that of CatL. Mutation of this loop into the CatL-like equivalent decreased elastin degradation by ∼70% and adding the Y118 mutation brought down the loss of elastolysis to ∼80%. In addition, the Y118 mutation selectively reduced the cleavage of the basement membrane component laminin by ∼50%. In summary, our data show that the degradation of elastin by CatS requires two exosites where one of them is distinct from those of CatK and V whilst the cleavage of laminin requires only one exosite.

Comparison of the efficacy of different techniques, culture media, and sources of blood in determining the hemolytic activity ofListeriaspp.

2001 ◽  
Vol 47 (7) ◽  
pp. 653-661 ◽  
Author(s):  
Rosa Capita ◽  
Carlos Alonso-Calleja ◽  
María Camino García-Fernández ◽  
Benito Moreno
Keyword(s):  
Blood Cell ◽  
Hemolytic Activity ◽  
Culture Media ◽  
Brain Heart Infusion ◽  
Listeria Innocua ◽  
Potassium Tellurite ◽  
Listeria Ivanovii ◽  
Hemolysis Test ◽  
Listeria Seeligeri ◽  
Inexpensive Procedure

Hemolytic activity is a fundamental criterion for the differentiation of Listeria species; therefore, a simple and inexpensive procedure to clearly distinguish hemolytic strains from each other and from nonhemolytic strains would be of great aid. We compared the efficacy of several techniques, culture media, and types of blood in demonstrating the hemolysis of Listeria spp. The hemolytic activities of Listeria monocytogenes and Listeria seeligeri were more easily detected with a red blood cell top-layer (RBCTL) technique and with a microplate technique than when the strains were streaked on blood agar (BA). Listeria ivanovii produced a marked hemolysis regardless of the technique employed. In general, the hemolytic activity of these three species was stronger on media containing brain heart infusion (BHI) agar and (or) potassium tellurite (PT). However, Listeria innocua produced questionable hemolytic reactions when nonselective culture media with BHI and PT were utilized, limiting the advantages gained by employing the two compounds. The RBCTL and the BA techniques disclosed greater hemolytic activity for L. monocytogenes, L. seeligeri, and L. ivanovii with sheep and guinea pig blood than with horse and human blood. When the microplate technique was used, all four kinds of blood were equally effective.Key words: Listeria spp., hemolysis, test comparison.

scholarly journals Enhancement of the behavioral effects of endogenous and exogenous cannabinoid agonists by phenylmethyl sulfonyl fluoride

2012 ◽  
Vol 62 (2) ◽  
pp. 1019-1027 ◽  
Author(s):  
R.E. Vann ◽  
D.M. Walentiny ◽  
J.J. Burston ◽  
K.M. Tobey ◽  
T.F. Gamage ◽  
...  
Keyword(s):  
Behavioral Effects ◽  
Phenylmethyl Sulfonyl Fluoride ◽  
Sulfonyl Fluoride ◽  
Cannabinoid Agonists

Comparison of the efficacy of different techniques, culture media, and sources of blood in determining the hemolytic activity of Listeria spp.

2001 ◽  
Vol 47 (7) ◽  
pp. 653-661 ◽  
Author(s):  
Rosa Capita ◽  
Carlos Alonso-Calleja ◽  
María Camino García-Fernández ◽  
Benito Moreno
Keyword(s):  
Hemolytic Activity ◽  
Culture Media

Identification and characterization of gelatin-cleavage activities in the apically located extracellular matrix of the sea urchin embryo

2000 ◽  
Vol 78 (4) ◽  
pp. 455-462 ◽  
Author(s):  
Justin Flood ◽  
Janice Mayne ◽  
John J Robinson
Keyword(s):  
Extracellular Matrix ◽  
Sea Urchin ◽  
Inhibitory Effect ◽  
Divalent Metal Ions ◽  
Gelatinase Activity ◽  
Sea Urchin Embryo ◽  
Phenylmethyl Sulfonyl Fluoride ◽  
Sulfonyl Fluoride ◽  
Identification And Characterization

We have identified and partially characterized several gelatinase activities associated with the sea urchin extraembryonic matrix, the hyaline layer. A previously identified 41-kDa collagenase/gelatinase activity was generally not found to be associated with isolated hyaline layers but was dissociated from the surface of 1-h-old embryos in the absence of Ca2+ and Mg2+. While hyaline layers, freshly prepared from 1-h-old embryos, were devoid of any associated gelatinase activities, upon storage at 4°C for 4 days, a number of gelatin-cleavage activities appeared. Comparative analysis of these activities with the 41-kDa collagenase/gelatinase revealed that all species were inhibited by ethylenediamine tetraacetic acid but were refractory to inhibition with the serine protease inhibitors, phenylmethyl sulfonyl fluoride and benzamidine. In contrast, the largely Zn2+ specific chelator 1,10-phenanthroline had markedly different effects on the gelatinase activities. While several of the storage-induced, hyaline-layer-associated gelatinase activities were inhibited, the 41-kDa collagenase/gelatinase was refractory to inhibition as was a second gelatinase species with an apparent molecular mass of 45 kDa. We also examined the effects of a series of divalent metal ions on the gelatin-cleavage activities. In both qualitative and quantitative assays, Ca2+ was the most effective activator while Mn2+, Cu2+, Cd2+, and Zn2+ were all inhibitory. In contrast, Mg2+ had a minimal inhibitory effect on storage-induced gelatinase activities but significantly inhibited the 41-kDa collagenase/gelatinase. These results identify several distinct gelatin-cleavage activities associated with the sea urchin extraembryonic hyaline layer and point to diversity in the biochemical nature of these species.Key words: gelatinase, sea urchin, extracellular matrix.

Sign in / Sign up

Export Citation Format

Share Document