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Processing Site (processing + site)

Distribution by Scientific Domains

Chemistry	50%
Earth and Environmental Science	33%

Selected Abstracts

Activation of hepatocyte growth factor activator zymogen (pro-HGFA) by human kallikrein 1-related peptidases

FEBS JOURNAL, Issue 5 2008
Shoichiro Mukai
Hepatocyte growth factor activator (HGFA) is a serine protease and a potent activator of prohepatocyte growth factor/scatter factor (pro-HGF/SF), a multifunctional growth factor that is critically involved in tissue morphogenesis, regeneration, and tumor progression. HGFA circulates as a zymogen (pro-HGFA) and is activated in response to tissue injury. Although thrombin is considered to be an activator of pro-HGFA, alternative pro-HGFA activation pathways in tumor microenvironments remain to be identified. In this study, we examined the effects of kallikrein 1-related peptidases (KLKs), a family of extracellular serine proteases, on the activation of pro-HGFA. Among the KLKs examined (KLK2, KLK3, KLK4 and KLK5), we identified KLK4 and KLK5 as novel activators of pro-HGFA. Using N-terminal sequencing, the cleavage site was identified as the normal processing site, Arg407,Ile408. The activation of pro-HGFA by KLK5 required a negatively charged substance such as dextran sulfate, whereas KLK4 could process pro-HGFA without dextran sulfate. KLK5 showed more efficient pro-HGFA processing than KLK4, and was expressed in 50% (13/25) of the tumor cell lines examined. HGFA processed by these KLKs efficiently activated pro-HGF/SF, and led to cellular scattering and invasion in vitro. The activities of both KLK4 and KLK5 were strongly inhibited by HGFA inhibitor type 1, an integral membrane Kunitz-type serine protease inhibitor that inhibits HGFA and other pro-HGF/SF-activating proteases. These data suggest that KLK4 and KLK5 mediate HGFA-induced activation of pro-HGF/SF within tumor tissue, which may thereafter trigger a series of events leading to tumor progression via the MET receptor. [source]

An experimental test of a visual-based push,pull strategy for control of wood boring phytosanitary pests

AGRICULTURAL AND FOREST ENTOMOLOGY, Issue 3 2009
Stephen Mark Pawson
Abstract 1,International phytosanitary standards require mandatory fumigation for key wood boring beetle pests prior to export. Pressure to reduce the use of toxic fumigants has created a need for alternative control techniques. 2,A visual based push,pull strategy that exploits a differential attraction to yellow and ultra violet (UV) lights was tested for its efficacy at controlling Cerambycidae. 3,The relative attraction of four ,push' lighting treatments [two yellow (high and low-pressure Sodium), one white (metal halide) and a control (no light)] to beetles was assessed. Highly attractive UV ,pull' traps were compared with a paired control trap, the difference used as a measure of the UV traps attractiveness to residual beetles attracted by ,push' lights. 4,Trap catch beneath the two yellow ,push' lights was more similar to the control (no light) treatment than the white light for both species. Control ,push' lights had the highest average catch of Arhopalus ferus, whereas white light was least attractive. This finding was counter intuitive to expectations, and potential mechanisms are discussed. The white ,push' light was most attractive to Prionoplus reticularis. 5,Ultraviolet ,pull' traps were highly attractive to residual beetles drawn to yellow ,push' light treatments. Relative attraction to the UV ,pull' traps beneath control and white ,push' lights differed between species. 6,The results obtained suggest that a push,pull strategy combining yellow site lighting with UV kill traps could provide site specific control of wood borers. Future research should attempt large-scale trials that are subject to competing alternative stimuli at a wood processing site. [source]

A novel prohormone processing site in Aplysia californica: the Leu,Leu rule

JOURNAL OF NEUROCHEMISTRY, Issue 6 2002
Amanda B. Hummon
Abstract Neuropeptides are a complex set of signaling molecules produced through enzymatic cleavages from longer prohormone sequences. The most common cleavage sites in prohormones are basic amino acid residues; however, processing is observed at non-basic sites. Cleavage at Leu,Leu sequences has been observed in three Aplysia californica prohormones. To further investigate this unusual event, native and non-native synthetic peptides containing Leu,Leu residues are incubated with homogenates of Aplysia californica ganglia and the resulting products monitored with MALDI MS. Cleavage near and between Leu,Leu residues is observed in the abdominal and buccal ganglia homogenates, confirming the presence of an unidentified peptidase. In addition, fractions from an HPLC separation of buccal ganglia homogenates also produce cleavages at Leu,Leu residues. Products resulting from cleavage at Leu,Leu sites are observed and are produced in larger amounts in acidic and neutral pH ranges, and cleavage is inhibited by the addition of EDTA, suggesting a metal is required for activity. [source]

Structural Investigation of the HIV-1 Envelope Glycoprotein gp160 Cleavage Site 3: Role of Site-Specific Mutations,

CHEMBIOCHEM, Issue 12 2004
Lucia Falcigno Dr.
Abstract Proteolytic processing of HIV gp160 to produce gp120 and gp41 is performed by PC enzymes. This process is a prerequisite for the virus infectivity, since both gp120 and gp41 participate in the virus HIV-1 entry mechanism. The structure of the gp120/gp41 junction remains to be elucidated, and the structural features required for molecular recognition between HIV-1 gp160 and proteolytic enzymes have not been clarified. Furin is the best PC candidate for the gp160 proteolytic processing known to date. In previous studies on model peptides, we have shown the relevance of an N-terminal helix for the proper recognition of the gp160 processing site by furin. Here we analyze the effect of point mutations in peptides lacking a regular N-terminal helix. To this end, we present the structure,activity characterization of three peptide analogues of the HIV gp160 processing site that all present mutations in proline at positions P3 and/or P2,, while sharing the same N-terminal sequence, containing helix-breaking D -amino acids. Conformational analysis of the peptides was carried out in solution by NMR techniques, and furin's efficiency in cleaving them was measured. Structural findings are presented and discussed in relation to the different exhibited activity. [source]

Phosphate regulates uranium(VI) toxicity to Lemna gibba L. G3

ENVIRONMENTAL TOXICOLOGY, Issue 1 2007
Martin Mkandawire
Abstract The influence of phosphate on the toxicity of uranium to Lemna gibba G3 was tested in semicontinuous culture with synthetic mine water developed as an analogue of surface water of two abandoned uranium mining and ore processing sites in Saxony, Germany. Six concentrations of uranium were investigated under five different supply regimes of PO43, at constant pH (7.0 ± 0.5) and alkalinity (7.0 ± 1.6 mg L,1 total CO32,). The results showed significant inhibition of specific growth rates in cultures exposed to the highest uranium concentrations (3500 and 7000 ,g U L,1) at lowest PO43, supply of 0.01 mg L,1. An increase of phosphate concentration from 0.01 to 8.0 mg L,1 resulted in an increase of EC50 from 0.9 ± 0.2 to 7.4 ± 1.9 mg L,1 (significant with Student's t test, P > 0.05). The accumulation of uranium in L. gibba increased exponentially with the increase in uranium concentration in cultures with 0.01 and 0.14 mg PO43, L,1. Accumulation also increased significantly when PO43, supply was increased from 0.14 to 1.36 mg PO43, L,1 for all uranium concentrations. However, as the supply of PO43, gradually increased from 1.36 to 8.0 mg PO43, L,1, uranium bioaccumulation increased slightly but insignificantly before leveling off. Uranium speciation modeling with PhreeqC geochemical code predicted increases in the proportions of uranyl phosphate species when PO43, concentrations increase in the media. Most of these uranyl phosphate species have a high probability of precipitation [saturation indices (SI) > 0.93]. Therefore, the alleviation of uranium toxicity to L. gibba with phosphates is due to interactions among components of the media, mainly uranyl and phosphate which results in precipitation. Consequently, bioavailable fractions of uranium to L. gibba are reduced. This might explain lack of consistent EC50 values for uranium to most aquatic organisms. © 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 9,16, 2007. [source]

Effect of sequence polymorphism and drug resistance on two HIV-1 Gag processing sites

FEBS JOURNAL, Issue 16 2002
Anita Fehér
The HIV-1 proteinase (PR) has proved to be a good target for antiretroviral therapy of AIDS, and various PR inhibitors are now in clinical use. However, there is a rapid selection of viral variants bearing mutations in the proteinase that are resistant to clinical inhibitors. Drug resistance also involves mutations of the nucleocapsid/p1 and p1/p6 cleavage sites of Gag, both in vitro and in vivo. Cleavages at these sites have been shown to be rate limiting steps for polyprotein processing and viral maturation. Furthermore, these sites show significant sequence polymorphism, which also may have an impact on virion infectivity. We have studied the hydrolysis of oligopeptides representing these cleavage sites with representative mutations found as natural variations or that arise as resistant mutations. Wild-type and five drug resistant PRs with mutations within or outside the substrate binding site were tested. While the natural variations showed either increased or decreased susceptibility of peptides toward the proteinases, the resistant mutations always had a beneficial effect on catalytic efficiency. Comparison of the specificity changes obtained for the various substrates suggested that the maximization of the van der Waals contacts between substrate and PR is the major determinant of specificity: the same effect is crucial for inhibitor potency. The natural nucleocapsid/p1 and p1/p6 sites do not appear to be optimized for rapid hydrolysis. Hence, mutation of these rate limiting cleavage sites can partly compensate for the reduced catalytic activity of drug resistant mutant HIV-1 proteinases. [source]