Top PDF Interaction of protein C inhibitor with the type II transmembrane serine protease enteropeptidase.

Interaction of protein C inhibitor with the type II transmembrane serine protease enteropeptidase.

Interaction of protein C inhibitor with the type II transmembrane serine protease enteropeptidase.

As mentioned above, native EP is a type II transmembrane serine protease. It contains an N-terminal hydrophobic segment from position 18 to 44, predicted to span the membrane [49,50]. The recombinant EP used is a mixture of two forms, in which the heavy chain is truncated and starts either at Leu41 or Ser118. N- terminal sequence analysis by Edman degradation revealed that also the bEK contains a mixture of two heavy chains starting at Gly53 and Ser118 respectively (personal communication: Dr. B. Sarg, Innsbruck Medical University, Austria). Phospholi- pids did not influence EP inhibition by PCI. Assuming a heparin- like bridging mechanism for the stimulatory effect of phospholipids on PCI-protease interactions, these results are not surprising, since it has been shown previously that a truncated EP lacking the transmembrane domain does not interact with phospholipid vesicles [51]. Supporting this data, a commercially available protein-lipid overlay assay containing membrane phospholipids was performed. We could not detect any binding of recombinant human EP to phospholipids (unpublished data). From our data it cannot be excluded that the interaction of PCI with the catalytically active light chain of EP is influenced by membrane anchoring of EP. However, the huge heavy chain lies in between the active center and the plasma membrane. This could hinder potential phospholipid-bridging of PCI and the light chain of EP. It is therefore not very likely that phospholipids involved in anchoring of EP could represent a bridge for bringing together PCI and EP.
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Identification and characterization of fusolisin, the Fusobacterium nucleatum autotransporter serine protease.

Identification and characterization of fusolisin, the Fusobacterium nucleatum autotransporter serine protease.

Fusobacterium nucleatum is an oral anaerobe associated with periodontal disease, adverse pregnancy outcomes and colorectal carcinoma. A serine endopeptidase of 61–65 kDa capable of damaging host tissue and of inactivating immune effectors was detected previously in F. nucleatum. Here we describe the identification of this serine protease, named fusolisin, in three oral F. nucleatum sub-species. Gel zymogram revealed fusobacterial proteolytic activity with molecular masses ranging from 55–101 kDa. All of the detected proteases were inhibited by the serine protease inhibitor PMSF. analysis revealed that all of the detected proteases are encoded by genes encoding an open reading frame (ORF) with a calculated mass of approximately 115 kDa. Bioinformatics analysis of the identified ORFs demonstrated that they consist of three domains characteristic of autotransporters of the type Va secretion system. Our results suggest that the F. nucleatum fusolisins are derived from a precursor of approximately 115 kDa. After crossing the cytoplasmic membrane and cleavage of the leader sequence, the C-terminal autotransporter domain of the remaining 96–113 kDa protein is embedded in the outer membrane and delivers the N-terminal S8 serine protease passenger domain to the outer cell surface. In most strains the N- terminal catalytic 55–65 kDa domain self cleaves and liberates itself from the autotransporter domain after its transfer across the outer cell membrane. In F. nucleatum ATCC 25586 this autocatalytic activity is less efficient resulting in a full length membrane-anchored serine protease. The mature serine protease was found to cleave after Thr, Gly, Ala and Leu residues at the P1 position. Growth of F. nucleatum in complex medium was inhibited when serine protease inhibitors were used. Additional experiments are needed to determine whether fusolisin might be used as a target for controlling fusobacterial infections.
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Cleavage specificity analysis of six type II transmembrane serine proteases (TTSPs) using PICS with proteome-derived peptide libraries.

Cleavage specificity analysis of six type II transmembrane serine proteases (TTSPs) using PICS with proteome-derived peptide libraries.

Based on phylogenetic analysis of the serine protease domains and the domain structure of the extracellular stem region, the TTSPs have been divided into four subfamilies (Figure 1A). The largest is the HAT/DESC subfamily, with currently 7 proteases: human/murine airway trypsin-like (HAT/MAT), differentially expressed in squamous cells carcinoma (DESC)1, and HAT-like 1–5 protease. HAT-like 2 and 3 are only expressed in rodents [12]. HAT is predominantly expressed in the trachea [13,14] whereas DESC1 is restricted to the epithelia of the skin and oral cavity [15,16]. TTSPs of the hepsin/TMPRSS/enteropeptidase subfam- ily include hepsin, mosaic serine protease large form (MSPL), type II transmembrane serine protease (TMPRSS) 2, 3, 4, 5 and enteropeptidase. These TTSPs are expressed predominantly in fetal liver and kidney [1], prostate [17] and on the brush-border of the duodenum, respectively [18]. The matriptase subfamily contains three highly homologous proteases; matriptase, matrip- tase-2, and matriptase-3, as well as a protein with an atypical mosaic structure, polyserase-1. Matriptase was originally identified from a human breast cancer cell line and shows the most ubiquitous pattern of expression of the TTSP family [9–11]. Matriptase-2 plays an important role in iron homeostasis [19–21] and is expressed primarily in the liver, where it suppresses hepcidin expression [22]. Matriptase-3 is expressed in brain, ovary, testis and salivary gland [23], whereas polyserase-1 is mainly restricted to skeletal muscle, liver, placenta and heart [24]. The fourth subfamily consists of a single TTSP, corin, which is mainly expressed in cardiomyocytes [25] and is involved in the activation of proatrial natriuretic peptide, a cardiac hormone that regulates blood pressure and cardiac function by promoting natriuresis, dieresis, and vasodilatation [26].
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Tryptogalinin is a tick Kunitz serine protease inhibitor with a unique intrinsic disorder.

Tryptogalinin is a tick Kunitz serine protease inhibitor with a unique intrinsic disorder.

Clade II in Figure 4A was the second best-supported Kunitz group that consisted of only scorpion venom peptides. These peptides present a unique Cys framework of either 6 or 8 Cys residues compared with other Kunitz peptides and they are all potent trypsin inhibitors that share a double Cys at the C-terminus (Figure S2). Analysis of the tertiary structure demonstrated that, compared with classical Kunitz, SdPI apparently adapts a new disulfide bridge at the C-terminus and lacks the archetypical Cys 2 and Cys 4 disulfide bridge [66]. The interaction site of SdPI at the N-terminus with trypsin, however, did not change due to this structural deviation. BmKTT-1 shows a 56% protein sequence identity with SdPI and thus demonstrates an ortholog protein form Figure 1. A multiple protein sequence alignment of the monolaris group from the I. scapularis sialome project [9]. The multiple sequence alignment is based within the Cys framework showing the conserved Cys residues (black) and the arrow points to the oddly placed Cys position that depicts a potentially ‘atypical’ Cys motif in a monolaris protein described in the sialome of this tick – the asterisk denotes the missing Cys residue. All the monolaris sequences are named after their GenBank accession number and the ‘atypical’ sequence (DN971582) is shown last (at the bottom of the sequence alignment).
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Functional analysis of a missense mutation in the serine protease inhibitor SPINT2 associated with congenital sodium diarrhea.

Functional analysis of a missense mutation in the serine protease inhibitor SPINT2 associated with congenital sodium diarrhea.

The mutation of the conserved Tyr163 in the 2 nd Kunitz domain (KD2) of HAI-2, as well as the corresponding Tyr mutation in the 1 st Kunitz domain (KD1) alone or together resulted in a loss of function of HAI-2 against all the intestinal serine proteases tested in our assay. The crystal structure of the 1 st Kunitz domain of the HAI-1 (HAI-1KD1) in complex with the catalytic domain of matriptase provides useful information at the atomic level on the possible mechanism underlying the loss of function of the HAI-2 Y163 or Y68C mutant [49]. The KD1 of HAI-1 structure adopts a pear-shape structure formed essentially by 2 loops that are stabilized by three disulfide bonds (figure 6). This binding mode of HAI-1 KD1 is common for the inhibition of other serine protease of Kunitz type [50]. The Tyr in KD1 conserved in HAI-1 and HAI-2 is in a close vicinity of cysteines (C259 and C283 in HAI-1) involved in a disulfide bond at the interface between the HAI-1 and matriptase (figure 6). By contrast to the Cys 259 and 283 of HAI-1 that interact with the catalytic triad of matriptase, the Tyr is surrounded by hydrophobic residues and does not participate directly to the interface between HAI-1 and matriptase. Simulation of the Tyr substitution by either Cys or Ser in the HAI1-KD1 that reproduces the Y68C or the Y68S in our experiments, resulted in a drop of the stability of the residues surrounding the Tyr by 2.8 kcal/mol and 3.3 kcal/mol (calculated with FoldX3.0) [51]. Such a decrease in the stability of the loop at the interface of the HAI-1 and matriptase does not seem to affect the binding of KDI with matriptase since in our experiments the Y68S HAI-2 mutant retains its inhibitory activity. Our exper- iments indicate that the Tyr to Cys substitution has a more profound effect on the conformation of the HAI-1 loops at the interface with matriptase. One interpretation is that the thiol group of the substituted Y68C or Y163C may bridge with either cysteines C47/C71 or C142/C166 of the HAI-2 loop normally involved in a disulfide bond that fits the protease binding site. Such aberrant disulfide bond is expected to disrupt the conformation of the Kunitz domain loops of HAI-2 and its interaction with the protease; this hypothesis is supported by our results showing that the Y68C and Y163C of the HAI-2 almost completely suppressed the inhibitory activity of HAI-2 on tmprss13 and prostasin. Interactions between HAI-2 and membrane-bound serine prote- ases are probably complex and not limited to the interaction between one Kunitz domain and the catalytic domain of the serine protease as shown by the crystal. In our experiments we expressed full-length HAI-2 (with 2 Kunitz domains) and serine proteases that may have several accessory domains potentially involved in protein-protein interaction.
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HATL5: a cell surface serine protease differentially expressed in epithelial cancers.

HATL5: a cell surface serine protease differentially expressed in epithelial cancers.

Cloning and Expression of Full-length Human HATL5 Human esophageal RNA was obtained from Biochain (Newark, CA). First strand cDNA synthesis was performed with Oligo (dT) primers using a RETROscript kit according to the manufacturers’ instructions (Ambion, Life Technologies, Grand Island, NY). Gene specific primers were designed for full-length human HATL5 using the deposited sequence for Homo sapiens transmembrane protease, serine 11B, mRNA, GenBank#BC126195.1. The primers 59- GCCACCATGTAC-AGGCACGGCATATC-39 and 59- GAGTCCAGTCTTGGATGTAATCC-39 were used to amplify the cDNA using a high-fidelity PlatinumHTaq polymerase (Invitrogen, Life Technologies, Grand Island, NY) which was then inserted into pcDNA 3.1/V5-His TOPOH TA (Invitrogen, Life Technologies, Grand Island, NY) in frame with a C-terminal HIS- tag and V-5 epitope. Constructs were verified by sequencing (ABI Prism 3730 DNA Analyzer, Invitrogen, Life Technologies, Grand Island, NY). Transfection of HEK293 and COS-7 cells (ATCC, Manassas, VA) was performed using Lipofectamine 2000 accord- ing to the manufacturer’s instructions (Invitrogen, Life Technol- ogies, Grand Island, NY). Cells were cultured in Dulbecco’s modified Eagle’s media (Gibco, Life Technologies, Grand Island, NY) supplemented with 10% fetal bovine serum (Atlanta Biologicals, Lawrenceville, GA). Transfection was performed with 4.0 mg full-length HATL5-containing plasmid DNA. Cells were lysed using RIPA buffer: 150 mM NaCl, 50 mM Tris/HCl, pH 7.4, 0.1% SDS, 1% NP-40, and protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO) and cleared by centrifugation at 12,0006g at 4Cu. Protein concentrations were determined using a Pierce BCA Protein Assay Kit (Thermo Scientific, Waltham, MA). Protein samples were separated by SDS-PAGE using 4–12% NuPAGE Bis-Tris gels (Life Technologies, Grand Island, NY) under reducing conditions and analyzed by western blotting using a primary V-5 mouse monoclonal antibody (Invitrogen, Life Technologies, Grand Island, NY) and a goat anti-mouse secondary alkaline phosphatase-conjugated antibody (Millipore, Billerica, MA).
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The effect of an HIV-1 viral protease inhibitor on staurosporine-induced apoptosis in immortalized mesangial cells

The effect of an HIV-1 viral protease inhibitor on staurosporine-induced apoptosis in immortalized mesangial cells

Whilst there are no caspase inhibitors that are currently available for therapeutic use, there are currently two classes of protease inhibitors that have been utilized in the treatment of some pathologies; the inhibitors of the HIV-1 viral protease and angiotensin- converting enzyme inhibitors. It is not known how these drugs may affect the caspase cascade and therefore whether they would be effective in preventing excessive apoptosis and thus glomerular cell loss in the late stages of progressive glomerular sclerosis.
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Xin Cui, Zheng-Rong Sun, Gao-Wei Ren, Gui-Li Wang, Ying Qi, Yan-Ping Ma and Qiang Ruan

Xin Cui, Zheng-Rong Sun, Gao-Wei Ren, Gui-Li Wang, Ying Qi, Yan-Ping Ma and Qiang Ruan

Betaherpesvirinae subfamily, is widely distributed in human populations. It can cause a minor or asymptomatic infec- tion in immunocompetent individuals, and cause severe disease in neonates and immunosuppressed individuals such as allograft transplant recipients and AIDS patients (1). Although the exact mechanisms are not known, a majority of researchers believe that a substantial portion of the HCMV-encoded proteins have the potential to affect virulence through cell tropism, immune evasion, molecular mimicry, or interference with host chemokines (2,3).
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Analysis stage 3: Finding patterns of interaction

Analysis stage 3: Finding patterns of interaction

When you are faced with analyzing two-way cross-classified experiments, you are almost always interested in whether the two factors interact or not, and if they do interact, combinations of the two factors are responsible for the interaction in the data. When there are no independent replications, there are no traditional tests for interaction. SAS® macros have been developed to provide user-friendly statistical software for the analysis and interpretation of interaction in two-way experiments. These newly developed macros are called the AMMI (Additive Main-effects and Multiplicative Interaction) macros. The AMMI models allow one to analyze two-way data with interaction even if there are no independent replications. In addition to fitting AMMI models and models such as Tukey’s single degree of freedom for nonadditivity Model, and Mandel’s bundle-of-straight-lines model, the macros provide many useful graphical displays including displays that help you determine the pattern of interaction when a pattern exists and that help you decide how many multiplicative interaction terms to include in the model. This paper describes how the AMMI macros can be installed, how they can be used, and the kinds of output they produce.
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Trypsin-like serine proteases in Lutzomyia longipalpis--expression, activity and possible modulation by Leishmania infantum chagasi.

Trypsin-like serine proteases in Lutzomyia longipalpis--expression, activity and possible modulation by Leishmania infantum chagasi.

Soluble protein samples were separated by 12% SDS-PAGE co- polymerized with protein substrates (gelatin, casein, hemoglobin and albumin) at 0.1% final concentration under non-reducing conditions [24]. Samples were loaded in the gel in non reducing buffer (125 mM Tris pH 6.8, 4% SDS, 20% glycerol and 0.002% bromphenol blue). Electrophoresis was carried out under constant voltage (100 V) at 4uC. The gels were washed in 2.5% Triton X-100 at 4uC under agitation during 1 hour to remove SDS. To test for best conditions, proteolysis of copolymerized proteins was performed by incubating the gel in different buffers: 0.1 M citrate (pH 4), or 0.1 M sodium phosphate (pH 6 and 8), or 0.2 M glycine (pH 10 and 12) for 18 hours at 37uC. All further zymography experiments were performed at pH 8. The proteolytic classes were determined by incubation of the gels at pH 8 buffer supplemented with the following specific protease inhibitors: trans-epoxysuccinyl L-leucylamido- (4-guanidino) butane (E-64) (10 m M) for cysteine-peptidases, pep- statin A (1 m M) for aspartic-peptidases, 1,10-phenanthroline (10 mM) for metallo-peptidases, phenyl-methyl sulfonyl-fluoride (PMSF) (1 mM) for serine-peptidases, tosyl phenylalanyl chloromethyl ketone (TPCK) (100 m M) for chymotrypsins, tosyl-lysyl-chloromethylketone (TLCK) (100 m M), benzamidine (1 mM), and 4-aminobenzamidine (1 mM) for trypsins. The gels were stained for 1 hour with 0.1% Coomassie blue R-250 in methanol:acetic acid:water (30:10:60) and destained in the same solvent.
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β-carboline compounds, including harmine, inhibit DYRK1A and tau phosphorylation at multiple Alzheimer's disease-related sites.

β-carboline compounds, including harmine, inhibit DYRK1A and tau phosphorylation at multiple Alzheimer's disease-related sites.

The dual-specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A) gene is located within the Down syndrome critical region on chromosome 21. Overexpression of DYRK1A has been proposed to be a significant contributor to the underlying neurodevelopmental abnormalities associated with Down syn- drome. Transgenic animals overexpressing DYRK1A show marked cognitive deficits and impairment in hippocampal dependent memory tasks [1,2]. Studies in cell culture models and transgenic models of Down syndrome that overexpress DYRK1A implicate the DYRK1A kinase in the generation of both amyloid and tau pathologies associated with the early onset Alzheimer’s disease (AD) that is uniformly observed in Down Syndrome [3,4,5,6]. We and others have shown that DYRK1A is important for phosphorylation of tau protein on multiple sites in several cellular models [3,4,6,7]. Interestingly, DYRK1A protein has been found to be associated with neurofibrillary tangles (NFTs) in sporadic Alzheimer’s disease [8] and some studies have found a genetic association between SNPs within the DYRK1A locus and
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N-terminal arginines modulate plasma-membrane localization of Kv7.1/KCNE1 channel complexes.

N-terminal arginines modulate plasma-membrane localization of Kv7.1/KCNE1 channel complexes.

complex. Positively-charged amino-acids may interact with hydrophilic phospholipid components of cellular membranes [16]. Arginine is typically located at membrane-water boundaries [17]. Several studies have identified a respective role for arginine in membrane localization. For instance, Jaskolski et al. described splice variants of a glutamate receptor where the variant containing a sequence of two arginines and two lysines showed significantly higher membrane localization [18]. Furthermore, membrane anchoring of saposin-C depends heavily on two lysines of the protein. Substitution of either lysine with neutral or negatively-charged amino-acids led to decreased anchoring while exchange with arginine intensified membrane anchoring [19]. Consistently, substitution of the three N-terminal KCNE1 arginines resulted in reduced current density and plasma- membrane localization in the present work. The reduction of membrane localization was comparable to N-terminal truncation of KCNE1 and glycine substitution suggesting a role of these amino-acids for KCNE1 function.
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In-silico patterning of vascular mesenchymal cells in three dimensions.

In-silico patterning of vascular mesenchymal cells in three dimensions.

The mathematical model admits up to 3 real uniform steady states for the parameter region we explored. Of these, one is always the zero solution{U ~0,V ~0,n = 1}, the other is low {U ~0:1,V ~0:2,n = 1}, and the third is high {U ~1:0,V ~3:0,n~1}. In the supplementary info, a linear stability analysis was carried out to analyze the stability of these steady states and determine the region where patterns are found. Briefly, the linear stability analysis analyzes a small perturbation from the steady state and determines which modes of the perturbation are unstable, which generally corresponds to the size of the perturbation. Among these states, the zero solution is always stable and the low solution is always unstable. The high state is stable with respect to spatially uniform perturbations, but it can be unstable with respect to spatially non-uniform modes. We performed simulations and analyzed the stability of these steady states (Supplementary Info) and found that only the higher steady state produced patterns that resembled the experiments and is likely the physiologically relevant one. We start with an initial condition at this steady state and add a 1% relative random noise to model cell variation[11]. The simulations shown in Figures 2 and 3 are the state distribution of cells with red color indicating high levels of cell density and blue levels indicating low levels of cell density. The lowest values of cell density are made transparent for visual clarity. The parameters used unless otherwise specified were D = 0.005, q = 0.003, x~10 { 5 , K = 0.25, B = 1.1, c = 600 and the box length
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The HtrA-like serine protease PepD interacts with and modulates the Mycobacterium tuberculosis 35-kDa antigen outer envelope protein.

The HtrA-like serine protease PepD interacts with and modulates the Mycobacterium tuberculosis 35-kDa antigen outer envelope protein.

Based on this data, we postulate that PepD functions to proteolytically regulate Rv2744c levels to help maintain cell wall/ cell envelope homeostasis in M. tuberculosis (Figure 4). A model is also proposed that builds upon observations previously reported by Barik et al [46] and others [3,47,48] concerning interactions between the SigE and MprAB signalling pathways in M. tuberculosis following exposure to extracytoplasmic stress. The serine/ threonine protein kinase, PknB, contains PASTA domains that have been postulated to bind peptidoglycan and may serve as cell wall sensors [49]. As the peptidoglycan becomes disordered due to extracellular stress, PknB activates and phosphorylates RseA, the anti-sigma factor of SigE. Phosporylation of RseA leads to proteolytic degradation of this protein by ClpC1P2, releasing SigE and inducing expression of components of the SigE regulon including mprA and clgR [39,46,50]. MprA and ClgR in turn upregulate gene products within their cognate regulons including clgR itself, clpC1, clpP2, ppk1, pepD, and sigE [3,34,35]. Upregula- tion of clp genes initiates a positive feedback loop through SigE by enhancing degradation of RseA. Similarly, upregulation of ppk1 encoding polyphosphate kinase increases polyphosphate levels and enhances activation of the MprAB two-component system [47], mediating a positive feedback loop through SigE [3]. The Rv2744c generated following upregulation of clgR is secreted extracytoplasmically, where it functions in an as-of-yet undefined role to help mediate resistance to the recognized stress. In Escherichia coli and other bacterial species, PspA forms higher order oligomers where the protein is thought to function as a structural scaffold to help maintain proton motive force [51,52,53,54]. While it is currently unclear if higher order oligomers are formed by Rv2744c in M. tuberculosis, Rv2744c can interact with itself in bacterial two-hybrid assays carried out in E. coli (Figure S2). Over- production of
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Effect of heat treatment parameters on the properties of low-alloy cast steel with microadditions of vanadium

Effect of heat treatment parameters on the properties of low-alloy cast steel with microadditions of vanadium

This article examines the effect of prolonged time of holding at the temperature of 620 0 C on the processes of secondary phase precipitation and mechanical properties of low-alloy cast steel with an addition of vanadium subjected to two variants of heat treatment, i.e. U:1150 0 C+H:950 0 C+O:620 0 C and H:950 0 C+O:620 0 C. To determine an impact of the applied heat treatment operations, testing of mechanical properties and microstructural examinations of the cast steel with 0,21 and 0,27%C were carried out.
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Aline Regiele PESOTI1 , Bruno Menezes de OLIVEIRA2 , Augusto Cesar de OLIVEIRA1 , Dávia Guimarães POMPEU1, Daniel Bonoto GONÇALVES

Aline Regiele PESOTI1 , Bruno Menezes de OLIVEIRA2 , Augusto Cesar de OLIVEIRA1 , Dávia Guimarães POMPEU1, Daniel Bonoto GONÇALVES

Tricine SDS-PAGE was carried out according to the procedure of Schägger & von Jagow (1987) with some modifications. The separating gel, spacer gel, and stacking gel were well prepared and solidified. For supernatant, the highest protein concentration sample was diluted to 1mg/mL. Precipitate samples were treated with the same method as supernatant. Then 0.5mL of diluted sample was mixed with 0.5mL of sample buffer [0.05M Tris-HCl at pH 6.8, 4% (w/v) SDS, 12% (v/v) glycerol, 0.01% (w/v) bromophenol blue, and 2% (v/v) 2-ME (or no for nonreducing)] effectively and heated for 3 min in a boiling water bath, then centrifuged at 10000 rpm for 3min. Then 10μL or 20μL of each sample was loaded into a well. Tricine SDS-PAGE was performed at 30V for 1h, and then up to 100V. After electrophoresis, the gels were stationed for 2h in stationary liquid [45.4% (v/v) methanol, 9.2% (v/ v) glacial acetic acid] and then stained for 2h using 0.1% (w/v) Coomassie Brilliant Blue R-250. After staining, the gels were distained using a 10% (v/v) acetic acid solution until a clear background was obtained. The proteins used as molecular weight standards for Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS–PAGE) were phosphorylase b (94kDa), bovine serum albumin (66kDa), ovalbumin (44kDa), bovine anhydrase carbonic (30kDa), inhibitor of trypsin (20kDa), lysozyme (14.kDa).
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Irreversible inhibitors of serine proteases based on the \03B2-lactam scaffold as potential drug candidates

Irreversible inhibitors of serine proteases based on the \03B2-lactam scaffold as potential drug candidates

At present, proteases are recognized not only as protein degrading enzymes, but also as important signalling molecules of a complex biochemical network, requiring tight regulation. 8 Disorders in protease regulation are known to be related with aetiology and progression of several pathologies. In particular, excessive proteolysis, which is most often a result of unwanted activation of protease signalling pathways, is related to tissue damage and/or aberrant processing of other proteins, playing a crucial role in cancer, cardiovascular, inflammatory, immunological, respiratory, neurodegenerative diseases, diabetes and infections. 9, 10 Thus, serine proteases, have emerged as important drug targets and one of the great challenges in medicinal chemistry is the design and development of serine protease inhibitors. 6, 8, 11 In Table 1.1 some known serine proteases targeted for pharmacological intervention are listed. Most of them have inhibitors currently under clinical trials. 9
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Network analyses reveal pervasive functional regulation between proteases in the human protease web.

Network analyses reveal pervasive functional regulation between proteases in the human protease web.

Protease biology is also complex due to the large protease numbers in humans (460) and mice (525), which form the second largest enzyme family after ubiquitin ligases in these organisms [24]. Moreover, an additional 93 and 103 are predicted to be inactive proteases in human and mouse, respectively, which often can function as dominant negative counterparts [24]. Protease numbers are almost equally distributed in the intracellular and extracellular environments, and other than some proteases that segue between these two compartments, this distribution partitions and limits their potential interactions with each other. In an effort to systematically comprehend this complex biology, proteases are grouped by the MEROPS database, which is assembled from biochemical experimental data curated from the literature, into seven classes, five of which are found in human and mouse, according to the active site residue catalyzing substrate cleavage, and into clans based on the structure of the active site [25]. Similarly, inhibitors are commonly grouped according to the class of proteases they inhibit, with several inhibitors exhibiting broad inhibitory activity against proteases from more than one class. Interactions between proteases of the same class are well established as part of classically described cascades of proteases such as the complement [26–28] and coagulation [29,30] systems, and newer recognized cascades such as kallikreins [31] and caspases in apoptosis [2,32–34]. However, wide-ranging additional protease interactions have also been proposed to extend more globally to link networks forming what was termed the protease web [35]. The protease web was defined as the universe of cleavage and inhibition interactions between proteases and their inhibitors. Stemming from examples in simple systems such as in vitro biochemical analyses and early in vitro and cell culture degradomics analyses of protease substrates [36–38], and mRNA transcript analyses in cancer upon administration of protease inhibitors or tissue inhibitor of metalloproteinase (TIMP) overex- pression and knockout studies [19], the protease web concept has been well supported. Extending terminomics analyses to in vivo Author Summary
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Rev. Saúde Pública  vol.36 número3

Rev. Saúde Pública vol.36 número3

Note - Social-economic condition of the residential city zone = considered: per capita income in minimum salaries + education level + water suply, inadequate drainage and garbage collection facilities The variable social-economic condition of the area where the patient lived was defined according to: the educational level of the head of the family (less than 4 years of schooling, 4 to 7 years of school education and 8 or more years of studies); the per capita income (up to half of the minimum salary, half to one, one to two, two to five, five to fifteen, fifteen to twenty and above twenty minimum salaries); proportion of residences with water supply; inadequate drainage system and residences without garbage collection [28]. For these variables, scores of 1 to 10 were attributed, added and classified as high, medium and low, in reference to the origin of the city zone of each patient. All the data were from the “Fundação Instituto Brasileiro de Geografia e Estatística (IBGE)”, 1991.
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White and gray solidification of the Fe-C eutectic

White and gray solidification of the Fe-C eutectic

and grey eutectics are equal. It can be thought that the system will try to choose the eutectic which has - the minimum undercooling (or the maximum growth rate), i.e. grey as long as the growth rate is less than V c and white for faster velocities. In

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