Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: 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)

ABSTRACT A 3.3-kb fragment of genomic DNA from bacteriophage Phi-Ea1h encoding an amylovoran-directed depolymerase lyase was sequenced, and three open reading frames (ORFs) were detected. The first ORF could encode a lysozyme and the second a holin that may form a pore supporting cell lysis by the lysozyme. The third ORF encodes a protein of 657 amino acids and deletion mutation in this DNA fragment abolished extracellular polysaccharide (EPS)-degrading activity. A putative promoter and a ribosome binding sequence were located in front of the gene. A polymerase chain reaction product spanning the gene was inserted into multi copy plasmids including fusions with a Histidine-tagged sequence to facilitate its purification on a nickel nitrilotriacetic acid column. Maximal activity of the purified protein was observed between pH 4 and 5 at 52 degrees C. Visualized by staining with fluorescein isothiocyanate-labeled lectin from Abrus precatorious, the enzyme degraded the EPS-capsules of Erwina amylovora. In virulence assays, no symptoms were detected for a low inoculum of an E. amylovora strain when pear slices were soaked in a solution of depolymerase in contrast to control slices without addition of the enzyme. Furthermore, gfp- or lux-labeled E. amylovora cells were not propagated, when their amylovoran capsules were removed by the depolymerase. The enzyme could be a tool for biological control of fire blight.
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PMID:Characterization of a Viral EPS-Depolymerase, a Potential Tool for Control of Fire Blight. 1894 30

The localization of sialic acids and antimicrobial substances in the foot pads of the cat was examined by lectin histochemical and immunohistochemical methods. The lectin binding patterns of the eccrine glands were suggestive of the existence of large concentrations of sialoglycoconjugates that terminated in Siaalpha2-3Gal1-4GlcNAc. Results were consistent with localization of O-linked (mucin-type) sialoglycoproteins with the Siaalpha2-6Gal/GalNAc sequence in the epidermal layers, especially the stratum spinosum. Additionally, antimicrobial peptides, such as lysozyme, secretory component, lactoferrin, and the peptide group of beta-defensins were demonstrated to be immunolocalised in the eccrine glandular cells. These substances, except for secretory component, were also distributed in the epidermal strata. The sialic acids and antimicrobial substances found in the eccrine glandular secretions and epidermis may play an essential role in the preservation of skin integrity in feline foot pads.
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PMID:Functional properties of feline foot pads as studied by lectin histochemical and immunohistochemical methods. 1909 95

The Chinese brown frog (Rana chensinensis) is a special amphibian in northern China, as it has been used widely in traditional Chinese medicine. The skin of the Chinese brown frog is also a promising resource for producing diverse antimicrobial peptides. To obtain a more comprehensive view of the metabolism and effective pharmacological components of Chinese brown frog skin, we constructed a non-normalized cDNA library from the skin. By sequencing cDNA clones at the 5' end, we obtained 5,976 high-quality EST sequences, which clustered into 512 contigs and 1379 singletons (in all 1,891 clusters). After BLAST searches of the protein and nucleic acid databases in GenBank, we found 46.7% of clusters to have significant similarity to known sequences; 28% matched Xenopus tropicalis ESTs and 29.1% matched Xenopus laevis ESTs. Gene annotation results indicated that genes related to secretion and defensive function, such as ubiquitin, lectin, and proteinase inhibitors, are highly expressed in the skin. Whey acidic-domain proteins are also highly abundant in the skin. Furthermore, both a beta-defensin and a lysozyme are transcribed in the frog skin, providing antibacterial protection. Analyses of gene ontology and KEGG metabolic pathways indicated the physiological roles of Chinese brown frog skin.
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PMID:Transcriptome analysis and identification of genes related to immune function in skin of the Chinese brown frog. 1926 15

Serum is an environment in which bacterial cells should not exist. The serum complement system provides innate defense against microbial infections. It consists of at least 35 proteins, mostly in pre-activated enzymatic forms. The activation of complement is achieved through three major pathways: the classical, alternative, and lectin. Lysozyme, widely present in body fluids, catalyzes the hydrolysis of beta 1,4 linkage between N-acetyloglucosamine and N-acetylmuramic acid in the bacterial cell wall and cooperates with the complement system in the bactericidal action of serum. In this study, ten strains of serotype O48 Salmonella, mainly associated with warm-blooded vertebrates and clinically important causing diarrhea in infants and children, were tested. The results demonstrated that the most efficient killing of Salmonella O48 occurred when all the components of normal bovine serum (NBS) and normal human serum (NHS) cooperated. To prove the role of lysozyme in the bactericidal activity of bovine and human serum, the method of serum adsorption onto bentonite (montmorillonite, MMT) was used. In order to investigate structural transitions accompanying the adsorption of serum components, we applied X-ray diffraction methods. The results of this investigation suggested that apart from lysozyme, other proteins (as, e.g., C3 protein or IgG immunoglobulin) were adsorbed on MMT particles. It was also shown that Ca(2+) cations can be adsorbed on bentonite. This may explain the different sensitivities of the serovars belonging to the same O48 Salmonella serotype to NBS and NHS devoid of lysozyme.
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PMID:Killing of Gram-negative bacteria with normal human serum and normal bovine serum: use of lysozyme and complement proteins in the death of Salmonella strains O48. 1929 63

The effects of some pathogen-associated molecular patterns (PAMPs) (laminarin, LPS and poly I:C) on total hemocyte counts (THC), phenoloxidase (PO) activity, superoxide anion production and lectin, prophenoloxidase, lysozyme, cytosolic manganese superoxide dismutase (C-MnSOD) and catalase (CAT) gene expression were studied. The results showed that the production or activity of most tested immune factors and the expression of most tested genes were up-regulated after stimulation with PAMPs, among which the highest value of lectin with 4.4 times as much as that of the control group appeared at 6 h in hemocytes, of CAT with 47 times as much as that of the control group appeared at 12 h in hepatopancreas, and with 2.7 times higher than that of the control group at 24 h of C-MnSOD in hepatopancreas after laminarin injection. The peak value of proPO, lysozyme and C-MnSOD appeared at 6 h in hepatopancreas, 24 h in hepatopancreas and 24 h in hemocytes after LPS injection, respectively. The highest expression level of lysozyme appeared at 12 h in hemocytes after poly I:C injection. However, significant decreases of PO activity in hemocytes and lectin expression in hepatopancreas were found after poly I:C injection, and a dramatic down-regulation of proPO expression from 3 h to 48 h was found in hemocytes after injection with laminarin, LPS and poly I:C. The results suggest that the shrimp immune response could be activated or inhibited by different PAMPs, and that the hepatopancreas also plays a key role by synthesizing immune factors.
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PMID:Immune response and gene expression in shrimp (Litopenaeus vannamei) hemocytes and hepatopancreas against some pathogen-associated molecular patterns. 1968 58

In this study, the immunostimulatory effect of oral administration of different preparations (conventional fine powder [CP] and superfine powder [SP]) of Astragalus membranaceus root or its polysaccharides (APS) in sea cucumber (Apostichopus japonicus) was investigated. Sea cucumbers with an average initial weight of 49.3 +/- 5.65 g were fed with a diet containing 3% CP or SP or 0.3% APS over a period of 60 days. The non-specific humoral (phenoloxidase, lysozyme and agglutination titer) and cellular (phagocytic capacity and reactive oxygen species) responses were determined and compared with controls (no supplement) after 20, 40 and 60 days of feeding. Variation in the levels of responses was evident among different supplements. SP and APS significantly enhanced most of the immune parameters tested. Among the humoral responses, lysozyme activity significantly increased after feeding with SP-supplemented diet for 20, 40 or 60 days. Furthermore, lectin titer showed significant enhancement after 20 and 60 days of feeding with APS-supplemented diet. Significant increase in the production of reactive oxygen species was evident for all three supplements after 20 days of feeding, but no significant change in serum phenoloxidase activity was observed for any of the three supplements over the three different periods. Overall, significant modulation of the cellular responses was only noticed after 20 days of feeding with SP- or APS-supplemented diet. After 60 days, these two groups also exhibited a decrease in the cumulative symptom rates compared to the controls when challenged with Vibrio splendidus. These results indicated that dietary intake containing A. membranaceus root or its polysaccharides could enhance the immune responses of A. japonicus and improve its resistance to infection by V. splendidus.
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PMID:Enhancement of non-specific immune response in sea cucumber (Apostichopus japonicus) by Astragalus membranaceus and its polysaccharides. 1974 88

3-Sulfo-N,N-dimethyl-N-(2'-methacryloyloxyethyl)propanaminium inner salt (SPB) was polymerized on a glass plate with a surface-confined initiator of atom transfer radical polymerization (ATRP) having a 2-bromoisobutyryl group. The glass plate modified with a brush of sulfobetaine telomer (PSPB) was highly hydrophilic and showed a strong resistance against nonspecific adsorption of proteins such as lysozyme and albumin. Through the polymerization from the free surface of PSPB chain by ATRP, furthermore, N-methacryloyloxysuccinimide (MAOSu) residues were introduced, and the incubation of the telomer (PSPB-b-PMAOSu)-modified glass chip with a lectin (concanavalin A, Con A) gave a glass chip covered with the Con-A-modified PSPB brush. The Con A fixed to the zwitterionic telomer brush pursued specific binding of mannose residues accumulated on the surface of Au colloidal particles, resulting in the increase in absorbance at 550 nm ascribable to localized surface plasmon resonance, while the nonspecific adsorption of proteins to the surface of the glass chip was still largely suppressed. The present results indicate usefulness of the zwitterionic telomer surface with antibiofouling properties as a scaffold for specific sensing devices.
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PMID:Molecular recognition at the exterior surface of a zwitterionic telomer brush. 2008 73

Due to the fact that some individuals are allergic to crustaceans, the presumed relationship between allergy and the presence of chitin in crustaceans has been investigated. In vivo, chitin is part of complex structures with other organic and inorganic compounds: in arthropods chitin is covalently linked to proteins and tanned by quinones, in fungi it is covalently linked to glucans, while in bacteria chitin is diversely combined according to Gram(+/-) classification. On the other hand, isolated, purified chitin is a plain polysaccharide that, at the nano level, presents itself as a highly associated structure, recently refined in terms of regularity, nature of bonds, crystallinity degree and unusual colloidal behavior. Chitins and modified chitins exert a number of beneficial actions, i.e., (i) they stimulate macrophages by interacting with receptors on the macrophage surface that mediate the internalization of chitin particles to be degraded by lysozyme and N-acetyl-beta-glucosaminidase (such as Nod-like, Toll-like, lectin, Dectin-1, leukotriene 134 and mannose receptors); (ii) the macrophages produce cytokines and other compounds that confer non-specific host resistance against bacterial and viral infections, and anti-tumor activity; (iii) chitin is a strong Th1 adjuvant that up-regulates Th1 immunity induced by heat-killed Mycobacterium bovis, while down- regulating Th2 immunity induced by mycobacterial protein; (iv) direct intranasal application of chitin microparticles into the lung was also able to significantly down-regulate allergic response to Dermatophagoids pteronyssinus and Aspergillus fumigatus in a murine model of allergy; (v) chitin microparticles had a beneficial effect in preventing and treating histopathologic changes in the airways of asthmatic mice; (vi) authors support the fact that chitin depresses the development of adaptive type 2 allergic responses. Since the expression of chitinases, chitrotriosidase and chitinase-like proteins is greatly amplified during many infections and diseases, the common feature of chitinase-like proteins and chitinase activity in all organisms appears to be the biochemical defense of the host. Unfortunately, conceptual and methodological errors are present in certain recent articles dealing with chitin and allergy, i.e., (1) omitted consideration of mammalian chitinase and/or chitotriosidase secretion, accompanied by inactive chitinase-like proteins, as an ancestral defensive means against invasion, capable to prevent the insurgence of allergy; (2) omitted consideration of the fact that the mammalian organism recognizes more promptly the secreted water soluble chitinase produced by a pathogen, rather than the insoluble and well protected chitin within the pathogen itself; (3) superficial and incomplete reports and investigations on chitin as an allergen, without mentioning the potent allergen from crustacean flesh, tropomyosine; (4) limited perception of the importance of the chemical/biochemical characteristics of the isolated chitin or chitosan for the replication of experiments and optimization of results; and (5) lack of interdisciplinarity. There is quite a large body of knowledge today on the use of chitosans as biomaterials, and more specifically as drug carriers for a variety of applications: the delivery routes being the same as those adopted for the immunological studies. Said articles, that devote attention to the safety and biocompatibility aspects, never reported intolerance or allergy in individuals and animals, even when the quantities of chitosan used in single experiments were quite large. Therefore, it is concluded that crab, shrimp, prawn and lobster chitins, as well as chitosans of all grades, once purified, should not be considered as "crustacean derivatives", because the isolation procedures have removed proteins, fats and other contaminants to such an extent as to allow them to be classified as chemicals regardless of their origin.
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PMID:Chitins and chitosans as immunoadjuvants and non-allergenic drug carriers. 2039 Jan 7

In this paper, the effect of temperature is investigated on the performance of glycoprotein enrichment by boronic acid lectin affinity chromatography (BLAC). Wheat germ agglutinin and m-aminophenyl boronic acid containing stationary phases were evaluated individually and in a mixed mode using an automated liquid handling robot with an integrated 96-well plate temperature controller. Glycoaffinity enrichment of the model proteins of ribonuclease B and trypsin inhibitor was investigated in the presence of the non-glycosylated proteins of myoglobin (neutral) and lysozyme (basic) at a wide temperature range of 5-65 degrees C. Our results revealed that glycoaffinity micropartitioning at the temperature of 25 degrees C provided the highest recovery rate for glycoprotein enrichment. We have also found that a large amount of lysozyme was present in the elution fractions of the m-aminophenyl boronic acid containing micropartitioning columns due to ion-exchange mechanism occurring between the positively charged protein and the negatively charged stationary phase at the operation pH. On the other hand, at high temperature (65 degrees C), non-specific interactions with the agarose carrier prevailed, evidenced by the presence of myoglobin in the eluate.
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PMID:Boronic acid lectin affinity chromatography (BLAC). 3. Temperature dependence of glycoprotein isolation and enrichment. 2049 Apr 66

A general rate model was employed in concert with a preferential interaction quadratic adsorption isotherm for the characterization of HIC resins and the prediction of solute behavior in these separation systems. The results indicate that both pore and surface diffusion play an important role in protein transport in HIC resins. The simulated and experimental solute profiles were compared for two model proteins, lysozyme and lectin, for both displacement and gradient modes of chromatography. Our results indicate that a modeling approach using the generate rate model and preferential interaction isotherm can accurately predict the shock layer response in both gradient and displacement chromatography in HIC systems. While pore and surface diffusion played a major role and were limiting steps for proteins, surface diffusion was seen to play less of a role for the displacer. The results demonstrate that this modeling approach can be employed to describe the behavior of these non-linear HIC systems, which may have implications for the development of more efficient preparative HIC separations.
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PMID:Characterization and modeling of nonlinear hydrophobic interaction chromatographic systems. 2125 85


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