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Query: EC:3.4.25.1 (
proteasome
)
28,817
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
In vivo most extracellular iron is bound to transferrin or lactoferrin in such a way as to be unable to catalyze the formation of hydroxyl radical from superoxide (.O2-) and hydrogen peroxide (H2O2). At sites of Pseudomonas aeruginosa infection bacterial and neutrophil products could possibly modify transferrin and/or lactoferrin forming catalytic iron complexes. To examine this possibility, diferrictransferrin and diferriclactoferrin which had been incubated with pseudomonas elastase, pseudomonas
alkaline protease
, human neutrophil elastase, trypsin, or the
myeloperoxidase
product HOCl were added to a hypoxanthine/xanthine oxidase .O2-/H2O2 generating system. Hydroxyl radical formation was only detected with pseudomonas elastase treated diferrictransferrin and, to a much lesser extent, diferriclactoferrin. This effect was enhanced by the combination of pseudomonas elastase with other proteases, most prominently neutrophil elastase. Addition of pseudomonas elastase-treated diferrictransferrin to stimulated neutrophils also resulted in hydroxyl radical generation. Incubation of pseudomonas elastase with transferrin which had been selectively iron loaded at either the NH2- or COOH-terminal binding site yielded iron chelates with similar efficacy for hydroxyl radical catalysis. Pseudomonas elastase and HOCl treatment also decreased the ability of apotransferrin to inhibit hydroxyl radical formation by a Fe-NTA supplemented hypoxanthine/xanthine oxidase system. However, apotransferrin could be protected from the effects of HOCl if bicarbonate anion was present during the incubation. Apolactoferrin inhibition of hydroxyl radical generation was unaffected by any of the four proteases or HOCl. Alteration of transferrin by enzymes and oxidants present at sites of pseudomonas and other bacterial infections may increase the potential for local hydroxyl radical generation thereby contributing to tissue injury.
...
PMID:Pseudomonas and neutrophil products modify transferrin and lactoferrin to create conditions that favor hydroxyl radical formation. 165 25
The authors propose a method for determination of proteolytic activity, based on the hydrolysis of
peroxidase
-labeled molecules of bovine serum albumin immobilized on the surface of polystyrene microassay plates with the subsequent determination of
peroxidase
activity on the carrier or in the solution. The optimum conditions for the sorption of the labeled substrate have been established. The method permits the determination of bacillary
alkaline protease
at a concentration of 01. microgram/ml within 45 minutes. The determination of four proteases has demonstrated that this method shows good correlation with the routine one (r = 0.98), but is more sensitive and less time- and labor-consuming.
...
PMID:[Method of determining proteolytic activity by using a conjugate of bovine serum albumin with peroxidase]. 245 31
Some regularities of the enzyme immunoassay (EIA) of whole bacterial cells have been studied on one of the bacillary species of contaminant microflora. Early detection of this microorganism is highly important for the microbiological production of alpha-amylase and
alkaline protease
(produced by Bacillus subtilis). The effective kinetic and equilibrant parameters of the interaction of
peroxidase
-labeled antibodies with the cells of the contaminant microflora in the solution and on the surface of the polystyrene plates have been defined. Two methods for the separation of cells after their interaction with
peroxidase
-labeled antibodies have been optimized: filtration involving the use of special filter plates and centrifugation in plates. The method for the immobilization of cells in the wells of standard assay plates by centrifugation has been proposed. Four EIA methods for measurement of contaminant microflora have been developed and optimized. These methods permit the determination of the microflora at concentration of 5 X 10(5)-5 X 10(4) cells/ml, depending on the scheme of the assay, within 1-3.5 minutes.
...
PMID:[Patterns in the immunoenzyme analysis of bacterial cells]. 312 Apr 44
The central projections of primary afferent fibers of the greater splanchnic nerve of the rat were investigated using the transganglionic horseradish
peroxidase
transport technique. In addition, the corresponding spinal ganglion cells and the preganglionic sympathetic neurons were demonstrated. For comparing visceral and somatic afferents, intercostal nerve afferents were labelled by the same technique. Splanchnic afferent dorsal root ganglion cells were found at segments T3 to T13 ipsilaterally, with the greatest density at T8 to T12. Labelled cells represented about 10%-15% of all neurons in the ganglia at maximal projection levels. They were randomly distributed within individual ganglia. The great majority were medium to small sized and round to slightly oval in shape. In the spinal cord, labelled visceral afferent axons were found maximally at T8 to T11, but could be detected in decreasing density up to T1 and down to L1. They were distributed over Lissauer's tract and the dorsal funiculus to a medial and lateral collateral pathway (
MCP
and LCP, respectively). The
MCP
, somewhat more prominent than the LCP, was destined primarily to clustered presumptive terminal fields in medial lamina I and outermost lamina IIa. Only a few axons continued further to laminae V and X. Splanchnic afferent axons, most likely derived from the
MCP
, formed a longitudinal bundle ventral to the central canal. The LCP consisted of more or less well-defined axon bundles emanating from the lateral Lissauer's tract and curving round the lateral edge of the dorsal horn and through the dorsolateral funiculus. Presumptive terminal sites of LCP axons are the lateral laminae I and IIa, the nucleus of the dorsolateral funiculus and the dorsal part of lamina V. A few LCP axons were seen in the vicinity of lateral dendrites of preganglionic sympathetic axons. Visceroafferent terminals were absent from laminae IIb-IV and VII. The possible consequences of the
MCP
/LCP duality for the central connections of splanchnic afferents are discussed. Some splanchnic afferents ascended to the gracile and cuneate nuclei, and rarely to the spinal trigeminal nucleus. These results fit into the general concept of visceroafferent terminal organization that has emerged during the last few years. Differences to other reports in the detailed arrangement of fibers and terminals are discussed. Somatoafferent cell bodies represented the vast majority of neurons in the respective spinal ganglia. Cell sizes encompassed the whole range from very small to very large without a clear predominance of one particular size class.(ABSTRACT TRUNCATED AT 400 WORDS)
...
PMID:The central projections of primary afferent neurons of greater splanchnic and intercostal nerves in the rat. A horseradish peroxidase study. 370 72
The inducible extracellular
alkaline protease
of Neurospora crassa was demonstrated to be a glycoprotein containing D-galactose residues by use of the enzyme-lectin conjugate horseradish
peroxidase
-Ricinus communis-agglutinin-120. The carbohydrate moiety of the protease appears to be a poor antigen since an antiserum made to the native enzyme recognizes epitopes determined only by the polypeptide portion of the enzyme. Immunochemical techniques were used to quantitatively precipitate protease labeled in vivo for electrophoretic analysis. Protease synthesis could not be detected in control, uninduced cultures, whereas ca. 0.4% of total cellular protein synthesis is devoted to protease formation under inducing conditions.
...
PMID:Major extracellular protease of Neurospora crassa. 623 9
The present study was designed to examine the effect of Pseudomonas aeruginosa
alkaline protease
and elastase on human polymorphonuclear leukocyte chemiluminescence. Both a luminol-enhanced and a nonenhanced chemiluminescence system using opsonized zymosan were utilized. It was found that
alkaline protease
and elastase at concentrations of 25 micrograms/ml strongly inhibited luminol-enhanced
myeloperoxidase
-mediated chemiluminescence, whereas inhibition of the nonenhanced chemiluminescence response was about 50%. In an attempt to determine the mechanism of inhibition of neutrophil chemiluminescence by these proteases, we examined the effect of various inhibitors of neutrophil oxidative metabolism on chemiluminescence, namely, superoxide dismutase, sodium azide, and catalase. It was shown that the pattern of inhibition of chemiluminescence by
alkaline protease
and elastase was similar to that of sodium azide, inhibitor of
myeloperoxidase
. The present study demonstrates that
alkaline protease
and elastase, extracellular products of P. aeruginosa, are capable of inhibiting
myeloperoxidase
-mediated chemiluminescence, one of the major antimicrobial systems of polymorphonuclear leukocytes. These findings provide further evidence for the role of P. aeruginosa exoproteases as virulence factors in the pathogenesis of infections caused by this microorganism.
...
PMID:Pseudomonas aeruginosa exoproteases inhibit human neutrophil chemiluminescence. 632 28
The central distribution of visceral primary afferent fibers from the pelvic nerve of the cast and the relationship of these fibers to preganglionic neurons of the sacral parasympathetic neurons (SPN) have been studied. Horseradish
peroxidase
(HRP) applied to the cut pelvic nerve was detected ipsilaterally in preganglionic neurons and dorsal root ganglion cells (segments S1-S3), and in central afferent projections to Lissauer's tract (LT), the dorsal columns, the dorsolateral funiculus, and spinal gray matter. The afferent projections were strongest in the region of the SPN (S1-S3) but extended far beyond its limits (e.g., LT was labeled from L4 to Cx7). In the transverse plane, collateral fiber bundles formed a thin shell around the dorsal horn predominantly within lamina I and expanded into terminal fields in the gray matter. The more prominent lateral collateral projection (LCP) extended into laminae V and VI, whereas the medial one (
MCP
) ended in the dorsal commissure. In longitudinal planes these projections exhibited a periodicity with an interval of approximately 200 micrometer. The distribution of afferent collateral projections overlaps the regions where many preganglionic neurons and their dendritic extensions are located, and also areas known to contain interneurons involved in visceral pathways. A differential distribution of afferents within the SPN was noted where a higher intensity was observed in proximity to those neurons located in laminae V and VI, which innervate the colon, and a lower intensity near neurons located in Lamina VII which innervate the bladder. This is consistent with the known spinal control of colon reflexes and the supraspinal control of bladder reflexes. The widespread rostrocaudal extent of the pelvic primary afferent projection is consistent with the necessity for the integration of somatic and autonomic elements from various levels of the lumbo-sacral-coccygeal spinal cord in the performance of pelvic visceral functions.
...
PMID:The distribution of visceral primary afferents from the pelvic nerve to Lissauer's tract and the spinal gray matter and its relationship to the sacral parasympathetic nucleus. 727 58
Significant progress has been made on the random sequencing of cDNAs (ESTs) and the genetic and physical mapping of the Arabidopsis thaliana genome. New techniques are now required to identify and map the expressed genes efficiently on A. thaliana chromosomes. A novel method to construct a transcription map of expressed genes or cDNAs in specific regions of the genome using DNA-latex particles has been developed. The region-specific DNA fragments prepared from six cosmid clones that constitute a contig covering the abi1 locus on chromosome 4 were covalently bound to latex particles. The DNA-latex particles were used for the selection of region-specific cDNAs. Sequence analysis of the cDNA clones revealed that ABI1, RPS2, casein kinase 1 (CK1), nucleosome assembly protein I (NAP) cDNAs and T20837 EST, which are situated within the contig near abi1 locus, were selected. These results indicate that the cDNAs in the specific region of the genome were faithfully selected with this method. Sequence analysis also indicated that 11 selected cDNAs were derived from novel genes located near the abi1 locus and that four of the selected cDNAs encode putative proteins that have sequence similarity to cationic
peroxidase
, phosphatidylserine decarboxylase 2 (PSD2), trans-caffeoyl CoA 3-O-methyltransferase (CCoAMT), and
proteasome
subunit XC3.
...
PMID:Rapid construction of a transcription map for a cosmid contig of Arabidopsis thaliana genome using a novel cDNA selection method. 930 Oct 97
The objectives of this study were to (1) assess the role of the 26S
proteasome
complex in regulating the expression of the inducible isoform of nitric oxide synthase (iNOS) and vascular cell adhesion molecule-1 (VCAM-1) in a model of chronic granulomatous colitis in vivo and (2) determine the role of the
proteasome
in regulating the inflammatory response observed in this model of chronic gut inflammation. The selective proteasome inhibitor MG-341 (0.3 mg/kg) was administered by gavage beginning immediately before the induction of colitis and continuing daily thereafter for the entire 14-day experimental period. We found that chronic
proteasome
inhibition using MG-341 significantly attenuated the peptidoglycan/polysaccharide (PG/PS)-induced up-regulation of iNOS in the colon and spleen and the consequent increase in plasma levels of nitrate and nitrite. Furthermore, we found that the proteasome inhibitor suppressed the up-regulation of the adhesion molecule VCAM-1 in the colon. We also found that MG-341 attenuated PG/PS-induced increases in macroscopic colonic inflammation, bowel wall thickness, colonic dry weight and colonic
MPO
activity. Treatment with MG-341 also significantly reduced PG/PS-induced increases in macroscopic spleen inflammation, spleen weight and spleen
MPO
activity. We conclude that the 26S
proteasome
complex plays an important role in regulating the PG/PS-induced up-regulation of iNOS and VCAM-1 in vivo and appears to be important in regulating colonic and splenic inflammation.
...
PMID:Proteasome inhibition attenuates nitric oxide synthase expression, VCAM-1 transcription and the development of chronic colitis. 931 79
The optimal level of oxygen-dependent microbicidal activity in human neutrophils depends on the generation of highly toxic products, including hypochlorous acid, by hydrogen peroxide in the presence of chloride anion and the neutrophil granule protein
myeloperoxidase
(
MPO
). The biosynthesis of
MPO
is normally restricted to the promyelocytic stage of myeloid development and includes N-linked glycosylation, heme insertion, proteolytic processing, subunit dimerization, and eventual targeting to the azurophilic granule. In the endoplasmic reticulum,
MPO
precursors interact transiently with calreticulin and calnexin, presumably in their capacity as molecular chaperones. In light of the important role of the
MPO
-H2O2-chloride system in human host defense, the relatively high prevalence of inherited
MPO
deficiency was an unanticipated insight provided by the widespread use of automated flow cytometry for the enumeration of leukocytes in clinical specimens. In many cases of inherited
MPO
deficiency, affected neutrophils have immunochemical evidence of precursor protein but lack the subunits of mature
MPO
,
peroxidase
activity, or the ability to chlorinate target proteins. To date, four genotypes have been reported to cause inherited
MPO
deficiency, each of which results in missense mutations. In the genotype Y173C, the mutant precursor is retained in the endoplasmic reticulum by virtue of its prolonged interaction with calnexin, and it eventually undergoes degradation in the 20S
proteasome
. In this way, the quality control system operating in the endoplasmic reticulum retrieves malfolded
MPO
precursors from the biosynthetic pathway and creates the biochemical phenotype of
MPO
deficiency. Thus
MPO
deficiency caused by Y173C joins the ranks of cystic fibrosis, protein C deficiency, and other genetic disorders that reflect abnormalities in protein folding.
...
PMID:Quality control in the endoplasmic reticulum: lessons from hereditary myeloperoxidase deficiency. 1048 5
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