277 research outputs found
DEVDase activity is induced in potato leaves during Phytophthora infestans infection.
Programmed cell death (PCD) occurs in plants, animals and several branches of unicellular eukaryotes as
a part of developmental and/or defense processes. Caspase proteases are universal mediators of animal
apoptosis, a type of PCD. In plants, there are not animal caspase homologs; therefore, the characterization
of caspase-like activities is of considerable importance to our understanding of PCD in plants. Here we
report for the first time the involvement of caspase-3-like activity in the resistance mechanism of potato
to Phytophthora infestans infection. We showed that disease development in infected potato leaves is
dependent of caspase-3-like activity. Unlike plant DEVDases previously reported, this DEVDase activity
was sensitive to the serine protease inhibitor PMSF. As reported for other subtilisin- like proteases with
caspase activity, potato DEVDase activity was mainly localized in the apoplast. We demonstrated that in
total protein extract DEVDase activity accounts for a 60% of serine pro
Antimicrobial activity of an aspartic protease from Salpichroa origanifolia fruits
Abstract: Plant proteases play a fundamental role in several processes like growth, development and in response to biotic and abiotic stress. In particular, aspartic proteases (AP) are expressed in different plant organs and have antimicrobial activity. Previously, we purified an AP from Salpichroa origanifolia fruits called salpichroin. The aim of this work was to determine the cytotoxic activity of this enzyme on selected plant and human pathogens. For this purpose, the growth of the selected pathogens was analysed after exposure to different concentrations of salpichroin. The results showed that the enzyme was capable of inhibiting Fusarium solani and Staphylococcus aureus in a dose-dependent manner. It was determined that 1·2 μmol l−1 of salpichroin was necessary to inhibit 50% of conidial germination, and the minimal bactericidal concentration was between 1·9 and 2·5 μmol l−1. Using SYTOX Green dye we were able to demonstrate that salpichroin cause membrane permeabilization. Moreover, the enzyme treated with its specific inhibitor pepstatin A did not lose its antibacterial activity. This finding demonstrates that the cytotoxic activity of salpichroin is due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of the AP could represent a potential alternative for the control of pathogens that affect humans or crops of economic interest. Significance and Impact of the Study: This study provides insights into the antimicrobial activity of an aspartic protease isolated from Salpichroa origanifolia fruits on plant and human pathogens. The proteinase inhibited Fusarium solani and Staphylococcus aureus in a dose-dependent manner due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of salpichroin suggests its potential applications as an important tool for the control of pathogenic micro-organisms affecting humans and crops of economic interest. Therefore, it would represent a new alternative to avoid the problems of environmental pollution and antimicrobial resistance.Fil: Díaz, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján; ArgentinaFil: Rocha, Gabriela Fernanda. Universidad Nacional de Luján; ArgentinaFil: Kise, Francisco. Universidad Nacional de Luján; ArgentinaFil: Rosso, A. M.. Universidad Nacional de Luján; ArgentinaFil: Guevara, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Parisi, M.G.. Universidad Nacional de Luján; Argentin
Apoplastic hydrophobic proteins involved in tuber defense response to P. Infestans
During infection, oomycetes secrete effectors into the plant apoplast where they interact with host resistance proteins. In response, large amounts of protease and protease inhibitors (PIs), are accumulated.
We analyzed differentially expressed Apoplastic Hydrophobic Proteins (AHPs) in potato tubers from Innovator (resistant) and Spunta (susceptible) cvs, after wounding and P. infestans infection.
Intercellular washing fluid was extracted from control, wounded or infected tubers at 0, 24 and 48 h and chromatographed into a PepRPCtmHR5/5 in FPLC. After elution with acetonitrile, fractions were analyzed by SDS-PAGE and proteins identified by MALDITOF- MS. Innovator cv. showed a higher basal AHP content and hydrophobicity than Spunta cv. In the latter, infection induced accumulation of patatins and PIs, whereas in Innovator cv. no changes in PIs accumulation were observed. In response to P.
infestans infection, lypoxigenase, enolase, annexin p34 and glutarredoxin/cyclophilin were accumulated in both cvs.
Hydrophobicity of AHPs was higher after 24 h of wounding and infection in both cultivars. These results suggest that an increase in AHPs concentration would be related with the protection against the oomycete and with the degree of resistance to pathogens. Finally, changes in hydrophobicity of Pis may induce changes in proteaseinhibitor interaction affecting the defense response
Cholesterol and membrane phospholipid compositions modulate the leakage capacity of the swaposin domain from a potato aspartic protease (StAPs-PSI).
Potato aspartic proteases (StAPs) and their swaposin domain (StAsp-PSI) are proteins with cytotoxic activity
which involves plasma membrane destabilization. The ability of these proteins to produce cell death varies
with the cellular type. Therefore, StAPs and StAsp-PSI selective cytotoxicity could be attributed to the different
membrane lipid compositions of target cells. In this work we investigate the possible mechanism by
which StAPs and StAsp-PSI produce selective membrane destabilization. Results obtained from leakage assays
show that StAsp-PSI is a potent inducer of the leakage of LUVs containing anionic phospholipids, especially
those containing phosphatidylglycerol. Based in these results, we suggest that the cytotoxic activity of
StAsp-PSI on pathogenic microorganisms could be mediated by the attraction between the exposed positive
domains of StAsp-PSI and the negatively charged microorganism membrane. On the other hand, our circular
dichroism spectroscopic measurements and analysis by size exclusion chromatography and followed by electrophoresis,
indicate that hydrophobic environment is necessary to StAsp-PSI oligomerization and both
StAsp-PSI disulfide bounds and membrane with negative charged phospholipids are required by StAsp-PSI
to produce membrane destabilization and then induce cell death in tumors and microorganism cell targets.
Additionally, we demonstrate that the presence of cholesterol into the LUV membranes strongly diminishes
the capacity of StAsp-PSI to produce leakag
Mathematical analysis of the two dimensional active exterior cloaking in the quasistatic regime
We design a device that generates fields canceling out a known probing field
inside a region to be cloaked while generating very small fields far away from
the device. The fields we consider satisfy the Laplace equation, but the
approach remains valid in the quasistatic regime in a homogeneous medium. We
start by relating the problem of designing an exterior cloak in the quasistatic
regime to the classic problem of approximating a harmonic function with
harmonic polynomials. An explicit polynomial solution to the problem was given
earlier in [Phys. Rev. Lett. 103 (2009), 073901]. Here we show convergence of
the device field to the field needed to perfectly cloak an object. The
convergence region limits the size of the cloaked region, and the size and
position of the device.Comment: submitted to Analysis and Mathematical Physic
Measurement of W Polarisation at LEP
The three different helicity states of W bosons produced in the reaction e+
e- -> W+ W- -> l nu q q~ at LEP are studied using leptonic and hadronic W
decays. Data at centre-of-mass energies \sqrt s = 183-209 GeV are used to
measure the polarisation of W bosons, and its dependence on the W boson
production angle. The fraction of longitudinally polarised W bosons is measured
to be 0.218 \pm 0.027 \pm 0.016 where the first uncertainty is statistical and
the second systematic, in agreement with the Standard Model expectation
Search for Anomalous Couplings in the Higgs Sector at LEP
Anomalous couplings of the Higgs boson are searched for through the processes
e^+ e^- -> H gamma, e^+ e^- -> e^+ e^- H and e^+ e^- -> HZ. The mass range 70
GeV < m_H < 190 GeV is explored using 602 pb^-1 of integrated luminosity
collected with the L3 detector at LEP at centre-of-mass energies
sqrt(s)=189-209 GeV. The Higgs decay channels H -> ffbar, H -> gamma gamma, H
-> Z\gamma and H -> WW^(*) are considered and no evidence is found for
anomalous Higgs production or decay. Limits on the anomalous couplings d, db,
Delta(g1z), Delta(kappa_gamma) and xi^2 are derived as well as limits on the H
-> gamma gamma and H -> Z gamma decay rates
Measurement of W Polarisation at LEP
The three different helicity states of W bosons produced in the reaction e+
e- -> W+ W- -> l nu q q~ at LEP are studied using leptonic and hadronic W
decays. Data at centre-of-mass energies \sqrt s = 183-209 GeV are used to
measure the polarisation of W bosons, and its dependence on the W boson
production angle. The fraction of longitudinally polarised W bosons is measured
to be 0.218 \pm 0.027 \pm 0.016 where the first uncertainty is statistical and
the second systematic, in agreement with the Standard Model expectation
Bose-Einstein Correlations of Neutral and Charged Pions in Hadronic Z Decays
Bose-Einstein correlations of both neutral and like-sign charged pion pairs
are measured in a sample of 2 million hadronic Z decays collected with the L3
detector at LEP. The analysis is performed in the four-momentum difference
range 300 MeV < Q < 2 GeV. The radius of the neutral pion source is found to be
smaller than that of charged pions. This result is in qualitative agreement
with the string fragmentation model
Z Boson Pair-Production at LEP
Events stemming from the pair-production of Z bosons in e^+e^- collisions are
studied using 217.4 pb^-1 of data collected with the L3 detector at
centre-of-mass energies from 200 GeV up to 209 GeV. The special case of events
with b quarks is also investigated.
Combining these events with those collected at lower centre-of-mass energies,
the Standard Model predictions for the production mechanism are verified. In
addition, limits are set on anomalous couplings of neutral gauge bosons and on
effects of extra space dimensions
- …