Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.17 (lysozyme)
 21,489 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

1. A cationic protease has been purified from the granule fraction of blood-donor leukocytes by a preparative method including precipitation by acetone and chromatography on Bio-Gel A 1.5 m, CM-Sephadex C-50 and Sephadex G-G-75. 2. The pH optimum against denatured bovine hemoglobin is 7.4. Gel chromatography indicated a molecular weight close to 23 000. 3. This neutral protease (EC 3.4.-.-) is able to split the synthetic esters Z-Ala-NPh and AcAla3OMe, its activity on the former substrate being 2.2 times greater than that of pancreatic elastase, on the latter the same. It differs crucially from pancreatic elastase in having small elastinolytic activity. 4. In cationic disk electrophoresis, neutral protease resolves into three protein bands with lower mobility than lysozyme: all bands exhibit esterolytic activity against 2-acetoxy-3-naphthoic acid o-toluidide, strongly suggesting that they represent isoenzymes. 5. The enzyme is completely inhibited by iPr2P-F, partially so by soybean trypsin inhibitor and Trasylol. Cysteine, EDTA and TosLysCH2Cl have no effect. 6. During chromatography on CM-Sephadex C-50 a more positively charged enzyme(s) was identified. This had hemoglobinolytic activity at pH 7.4 but only a small esterolytic effect on Z-Ala-NPh; it showed only traces of activity against AcAla3OMe.
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PMID:Purification and some properties of a neutral protease from human leukocyte granules and its comparison with pancreatic elastase. 0 9

When lysozyme is reacted with 4-chloro-7-nitrobenz-2-oxa-1,3-diazole (NBD-CL), A 1:1 covalent product is produced, in which the NBD group arylates the phenolic hydroxyl group of Tyr-23 (Aboderin, A. A., and Boedefeld, E. (1976) Biochim. Biophys. Acta 420, 177). Changing the pH from neutral to alkaline conditions results in a large spectral shift of the absorption band associated with the NBD chromophore (Aboderin, A. A., Boedefeld, E., and Luisi, P. L. (1973) Biochim. Biophys. Acta 328, 30). In the present work it is shown that this spectral change is due to the formation of a sigma complex in which a hydroxyl ion is added to the aromatic nucleus of the nitrobenzoxadiazole system. Circular dichroic studies suggest that the NBD group is held in a conformationally rigid state in the protein. The kinetics of the spectral change accompanying the formation of the sigma complex has been investigated with a rapid mixing stopped flow spectrophotometer both in the modified enzyme and in the low molecular weight model compounds N-acetyl-(O-NBD)-L-tyrosinamide and glycyl-(O-NBD)-L-tyrosine. In the pH range from 10.1 to 12.7, the time course of the reaction is first order in the case of the modified enzyme (k = 4.8 s-1) and bimolecular and much slower (under pseudo-first order conditions) in the low molecular weight compounds. It is suggested that in the enzyme the reaction proceeds much faster because of the hydrophobic environment around the reacting groups. It is further suggested that the unimolecularity in the enzyme is due to a rate-determining isomerization step, probably connected with a local rearrangement of the protein conformation following the ionization of Tyr-20.
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PMID:Alkaline structural transition of 4-nitrobenz-2-oxa-1,3-diazolyl-Lysozyme. Kinetic and spectroscopic investigations. 1 90

The prevalence of respiratory diseases was followed in a group of 80 children (39 boys and 41 girls) of mean age 12.4 +/- 0.9 years during the part of school year (November - May incl.). The specimen of blood was taken from all studied children before the study started and the values of nine serum proteins were measured (IgG, IgA, IgM, IgE, alfa 1-antitrypsin = A 1-AT, transferrin = TRF, ceruloplasmin = CPL, alpha 2-macroglobulin = A 2M, lysozyme = LYS), as well as levels of two secretory proteins in saliva (sIgA and sLYS). The difference between the subgroup of boys, who fall into some respiratory illness during the observation time, and those who remain healthy, did not reach the level of significance. In contrast, the girls subgroup revealed significantly higher respiratory morbidity. With those girls, who remained healthy, the levels of serum IgE were highly increased and A 1-AT level weakly increased compared to ill girls. The level of TRF and CPL were significantly decreased with the healthy girls.
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PMID:The interrelations of some humoral factors of resistance with the prevalence of respiratory diseases in children. I. The differences among the protein levels in healthy and ill children. 617 77

Blood and saliva were collected in the autumn and spring from a group of schoolchildren (39 girls, 35 boys) with a mean age of 11.4 years. Serum immunoglobulin IgG, IgA, IgM and IgE, alpha 1-antitrypsin (A 1-AT), alpha 2 macroglobulin (A 2M), transferrin (TRF), ceruloplasmin (CPL), lysozyme (LYS) and pertussis (PE) antibody levels were determined. Calcium (Ca2+) and total serum protein levels were also determined. Secretory IgA (sIgA) and secretory lysozyme (sLYS) levels were assessed in the saliva. A highly significant drop in Ca2+ levels was found in the spring in boys, while in girls there was only a greater scatter of the values. Mean IgG, IgA and IgM values fell significantly in the spring in both sexes, but IgE levels fell significantly only in boys. PE levels rose significantly in the spring in girls. Among the other proteins, all the values rose in boys, except for TRF, whose levels fell. In girls, LYS and TRF levels rose, but all the other values fell. The coefficients of correlation between Ca2+ and the tested proteins showed a significant relationship only for A 2M and PE in girls and only for the total protein level in boys; in boys, the determination coefficient for sIgA and IgM was over 10%. The results do not testify to the existence of a close relationship between blood Ca2+ levels and Ig and other blood protein levels.
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PMID:Seasonal changes in the relationship of blood calcium levels to immunoglobulins and some of the blood proteins in schoolchildren. 650 75

The genus Rhodococcus comprises aerobic gram-positive actinomycetes that show considerable morphological and metabolic diversity and are known to be involved in the development of plant diseases and degradation of environmental pollutants. We describe a method for cell lysis and large plasmid DNA isolation from Rhodococcus by creating lysozyme susceptible cells by predigestion with the enzyme mutanolysin. Mutanolysin action resulted in the liberation of reducing sugars and free amino acids from the peptidoglycan layers of the cell wall. A 1-h predigestion with mutanolysin followed by a 0.5-h incubation with lysozyme resulted in spheroplast formation. Complete lysis of cells and efficient isolation of intact large plasmid DNA (108 kb) from wild-type Rhodococcus strains was confirmed.
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PMID:Spheroplast formation and plasmid isolation from Rhodococcus spp. 829 32

A metalloendopeptidase (MEP) isolated from rabbit liver microsomes with substrate specificity for peptides containing Arg at the P1 and P4 positions has recently proved to be identical to soluble angiotensin-binding protein present in the cytosol. Here we describe the peptide-degrading specificity of MEP, determined using various bioactive peptides and novel fluorogenic substrates for the enzyme. MEP degraded oligopeptides, including bradykinin, alpha-neoendorphin, bovine adrenal medulla dodecapeptide, substance P, bombesin, neurotensin, and alpha-endorphin, but not polypeptides such as reduced lysozyme and histone H4, hence, MEP probably belongs to the family of endo-oligopeptidases. It cleaved most preferentially at the -Phe-Ser- bond of bradykinin (kcat/Km = 2.8 x 10(4) M-1.S-1) but did not cleave high molecular weight and low molecular weight kininogens, the precursors of bradykinin. MEP did not cleave angiotensin I, dynorphin A 1-13, somatostatin, and luteinizing hormone-releasing hormone, some of which are good substrates for metalloendopeptidase-24.15, metalloendopeptidase-24.16, N-arginine dibasic convertase, and yeast endopeptidase-24.15 related peptidase. An active site-directed inhibitor of metalloendopeptidase-24.15, N-[1-(R,S)-carboxyl-3-phenylpropyl]-Ala-Ala-Phe-p-aminobenzoate also had no effects on the amidolytic activity of MEP. Based on the cleavage sites of bioactive peptides and processing sites of vitamin K-dependent proproteins, intramolecularly quenched fluorogenic peptide substrates were newly synthesized. Among the thirteen substrates used, the most reactive was 2-aminobenzoyl-Ala-Arg-Val-Arg-Arg-Ala- Asn-Ser-2,4-dinitroanilinoethylamide (kcat/Km = 9.3 x 10(5) M-1.S-1). An angiotensin antagonist, [Sar1, Ala8]-angiotensin II, inhibited hydrolysis of the substrate by MEP in a competitive manner (Kl = 7.6 microM). MEP cleaved oligopeptides even on the carboxyl side of proline residue and these peptides are resistant to hydrolysis by the cytosol-derived proteasome, therefore MEP may participate in the catabolism of oligopeptides in the cytosol, together with other endo-oligopeptidases.
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PMID:Substrate specificity of rabbit liver metalloendopeptidase and its new fluorogenic peptide substrates. 857 4

A 1.4-kb gene encoding the "small" sialidase isoenzyme of Clostridium perfringens A99, including its own promoter, was previously cloned in and expressed by Escherichia coli JM 101. Since all attempts to purify this enzyme to homogeneity were unsuccessful, a new strategy was developed. The structural gene was amplified by means of a PCR technique and inserted into the plasmid vector pQE-10, transferring a six-histidine affinity tag (His6) to the N-terminus of the protein. In order to minimize proteolytic degradation of the sialidase protein, the gene was subcloned into the Escherichia coli strain BL21(DE3)pLys S with reduced protease activity. The sialidase production was increased about 2.5-fold when compared with that of the original clone. The enzyme, released by lysozyme treatment of the bacterial cells, was purified by metal chelate chromatography on Ni-nitrilo-triacetic acid agarose to apparent homogeneity in SDS-PAGE. The 42-kDa protein was enriched 62-fold with a yield of 82% and a specific activity of 280 U mg-1. A total amount of 1 mg sialidase was obtained from 1 liter of bacterial culture. For future studies, including crystallization experiments, the histidine affinity tag was removed from the sialidase enzyme by aminopeptidase K. The sialidase was then separated from aminopeptidase K by ion-exchange chromatography, resulting in an overall yield of 83% and a specific activity of 305 U mg-1 using 4-methylumbelliferyl- alpha-D-N-acetylneuraminic acid under standard conditions. The two forms (with or without the histidine tag) of sialidase exhibited similar kinetic properties when compared to the wild-type enzyme.
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PMID:Expression and purification of a recombinant "small" sialidase from Clostridium perfringens A99. 877 61

Adhesion of yeasts and bacteria to silicone rubber is one of the first steps in the biodeterioration of silicone rubber voice prostheses. In this paper, adhesion of two streptococcal, staphylococcal, Candida albicans and Candida tropicalis strains, isolated from explanted voice prostheses was investigated to silicone rubber with and without a salivary conditioning film in a parallel-plate flow chamber. Within each microbial pair of one species, the strain with the most negative zeta potential adhered most slowly to negatively charged silicone rubber. No other clear relationships were obvious between adhesion to silicone rubber and microbial zeta potentials of cell-surface hydrophobicities, as by water contact angles. A 1.5-h adsorbed salivary conditioning film appeared to possess components, presumably albumin and lysozyme, slowing down the deposition of the yeasts and some of the streptococcal and staphylococcal isolates. In addition, microbial adhesion in a stationary end point was generally lower to silicone rubber with an adsorbed salivary conditioning film than without one. Nearly all microorganisms adhering to an adsorbed salivary conditioning film, yeasts as well as bacteria, were stimulated to detach by the passage of an air bubble through the chamber, but microorganisms adhering directly to the silicone rubber, especially C tropicalis strains, detached in far lower numbers under the influence of a passing air bubble. The present observations are in agreement with clinical in vivo findings that in patients with reduced saliva production after radiotherapy, the device life of the voice prosthesis is significantly shortened and suggests that isolated salivary components might be used as an anti-adhesive.
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PMID:Adhesion to silicone rubber of yeasts and bacteria isolated from voice prostheses: influence of salivary conditioning films. 902

The aim of this study was to further investigate effects of a combined chitosan and collagen matrix on osteogenic differentiation of rat-bone-marrow stromal cells (BMSCs), including analysis of the physical and mechanical properties of the sponges. There were 4 study groups: collagen, chitosan, 1:1 chitosan-collagen and 1:2 chitosan-collagen sponges. Chitosan-collagen sponges were fabricated using the freeze-drying technique. BMSCs were seeded on the sponges and cultivated in mineralized culture medium for 27 days. Attachment and growth of cells on the sponges were examined under a scanning electron microscope. Alkaline phosphatase activity and levels of osteocalcin were monitored. Tests of swelling, collagenase and lysozyme enzymatic degradation, and mechanical strength were performed. The BMSCs attached successfully to the structure of the sponges, and expression of ALP and osteocalcin on collagen and chitosan-collagen composite sponges was greater than on chitosan sponges. All sponges showed a high degree of water uptake. Chitosan and chitosan-collagen sponges showed a higher resistance to enzymatic degradation than collagen sponges. A 1:1 chitosan-collagen sponge demonstrated the highest compressive strength. Combined chitosan-collagen matrixes promoted osteoblastic differentiation of BMSCs, and improved the mechanical and physical properties of the sponges.
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PMID:Properties of chitosan-collagen sponges and osteogenic differentiation of rat-bone-marrow stromal cells. 1827 41

A 1-year-old intact female miniature Dachshund was presented with hematochezia, vomiting, and diarrhea of more than 1-week duration. An abdominal mass was palpated, which at exploratory surgery was found to be a 7-cm-long thickened section of ileum. The thickened ileum was resected. Impression smears revealed numerous small- to medium-sized lymphocytes, with a smaller number of cells resembling Mott cells. The Mott-like cells contained multiple pale vacuoles that were positive for periodic acid-Schiff (PAS) in wet-fixed smears, consistent with Russell bodies. Histologic evaluation of the surgically excised ileum revealed 2 populations of neoplastic lymphoid cells. The majority were uniform medium-sized lymphocytes with hyperchromatic oval or round nuclei and inconspicuous nucleoli. The remaining cells resembled Mott cells, which contained several PAS-positive eosinophilic globules in the cytoplasm, occasionally compressing the nucleus. The majority of neoplastic cells stained positively for vimentin, CD20, CD79a, and Pax-5, but were negative for CD3 and lysozyme; 43.5% of cells stained positively for Ki-67. The Mott cells were strongly positive for immunoglobulin but were negative for Pax-5. Using electron microscopy, a homogenous substance of intermediate electron density was observed frequently in the cisternae of rough endoplasmic reticulum in the cytoplasm of the Mott cells, and rarely in the perinuclear cisternae of the lymphoid cells, corresponding to the site of immunoglobulin staining. Monoclonal rearrangement of immunoglobulin heavy-chain (IgH) gene was observed by PCR testing for lymphocyte-antigen receptor rearrangement. The morphologic features, immunophenotype, and IgH gene rearrangement verified the lymphoid cells were neoplastic (mature cell type) and had a B-cell phenotype, with evidence of immunoglobulin production and differentiation into Mott cells. This case was unusual because of the age of the dog and because most intestinal lymphomas are T-cell phenotype. The Mott cell morphology also differed from typical mature B-cell lymphoma types and may be a unique B-cell lymphoma variant.
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PMID:B-cell intestinal lymphoma with Mott cell differentiation in a 1-year-old miniature Dachshund. 1905 69


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