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1.
The dependence of cold agglutinin activity on K chains 总被引:1,自引:0,他引:1
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Cauerhff A Braden BC Carvalho JG Aparicio R Polikarpov I Leoni J Goldbaum FA 《Journal of immunology (Baltimore, Md. : 1950)》2000,165(11):6422-6428
Cold agglutinins (CAs) are IgM autoantibodies characterized by their ability to agglutinate in vitro RBC at low temperatures. These autoantibodies cause hemolytic anemia in patients with CA disease. Many diverse Ags are recognized by CAs, most frequently those belonging to the I/i system. These are oligosaccharides composed of repeated units of N:-acetyllactosamine, expressed on RBC. The three-dimensional structure of the Fab of KAU, a human monoclonal IgM CA with anti-I activity, was determined. The KAU combining site shows an extended cavity and a neighboring pocket. Residues from the hypervariable loops V(H)CDR3, V(L)CDR1, and V(L)CDR3 form the cavity, whereas the small pocket is defined essentially by residues from the hypervariable loops V(H)CDR1 and V(H)CDR2. This fact could explain the V(H)4-34 germline gene restriction among CA. The KAU combining site topography is consistent with one that binds a polysaccharide. The combining site overall dimensions are 15 A wide and 24 A long. Conservation of key binding site residues among anti-I/i CAs indicates that this is a common feature of this family of autoantibodies. We also describe the first high resolution structure of the human IgM C(H)1:C(L) domain. The structural analysis shows that the C(H)1-C(L) interface is mainly conserved during the isotype switch process from IgM to IgG1. 相似文献
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The new enzyme phenylalanine dehydrogenase [L-phenylalanine: NAD+-oxidoreductase (deaminating)] was detected in the crude extract of a strain of Brevibacterium spec. The bacterium was isolated from a soil sample by enrichment with phenylalanine. This strain was the only one containing phenylalanine dehydrogenase out of 173 tested strains, among them 22 of the genus Brevibacterium, 74 strains from soil samples and 77 strains from a culture collection belonging to several genera. The enzyme is involved in the degradation of phenylalanine and could be induced by addition of L-, D-, D,l-phenylalanine or L-histidine, the optimum inducer concentration of phenylalanine being 1%.The reaction mechanism of a reductive amination was confirmed by demonstrating the close coupling between NADH-consumption and phenylalanine production; ammonia could not be replaced by L-glutamate or L-aspartate as amino donor. The -keto acid of L-tyrosine was converted too, while the corresponding compound of histidine was inactive. The optimum pH value for reductive amination in the crude extract was 8.5 and for oxidative desamination 10.5. 相似文献
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M Monsigny C Sene A Obrenovitch A C Roche F Delmotte E Boschetti 《European journal of biochemistry》1979,98(1):39-45
The physicochemical and binding properties of succinylated wheat germ agglutinin are described in comparison with these of unmodified wheat germ agglutinin. Succinylated wheat germ agglutinin is an acidic protein with a pI of 4.0 +/- 0.2 while the native lectin is basic, pI of 8.5. The solubility of succinylated wheat germ agglutinin is about 100 times higher than that of the unmodified lectin at neutral pH. Both lectins are dimeric at pH down to 5, and the dissociation occurs at pH lower than 4.5. The binding of oligosaccharides of N-acetylglucosamine to both lectins is very similar on the basis of fluorescence and phosphorescence studies. The minimal concentration required to agglutinate rabbit red blood cells is about 2 microgram/ml with both lectins and the concentrations of N-acetylglucosamine and di-N-acetylchitobiose which inhibit agglutination are similar with both lectins. The number of succinylated wheat germ agglutinin molecules bound to the surface of mouse thymocytes was ten times lower than that of the unmodified lectin although the apparent binding constant was only slightly different between the two lectins. The dramatic decrease of the apparent number of cell surface receptors upon succinylation of the lectin is discussed on the basis of the decrease of the isoelectric point and of the acidic properties of the cell surface. 相似文献
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C M Tsai D A Zopf V Ginsburg 《Biochemical and biophysical research communications》1978,80(4):905-910
Cold agglutinin MKV is a Waldenström macroglobulin that agglutinates human erythrocytes in the cold by binding -acetylneuraminosyl-containing carbohydrate chains on their surfaces. Neuraminidase-treated cells are not agglutinated but their reactivity can be restored by allowing them to adsorb hematoside (NeuNAcα2-3Galβ1-4Glcβ1-ceramide). When between 7 × 104 and 106 molecules are adsorbed per cell, the cells are agglutinated at 0° but not at 37°. When over 106 molecules of hematoside are adsorbed, they are agglutinated at both 0° and 37°. The density of receptors on the erythrocyte surface can thus influence the thermal amplitude of cold agglutinins. 相似文献
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《Bioorganic & medicinal chemistry letters》2014,24(22):5234-5237
The abietane-type diterpenoid (+)-ferruginol, a bioactive compound isolated from New Zealand’s Miro tree (Podocarpus ferruginea), displays relevant pharmacological properties, including antimicrobial, cardioprotective, anti-oxidative, anti-plasmodial, leishmanicidal, anti-ulcerogenic, anti-inflammatory and anticancer. Herein, we demonstrate that ferruginol (1) and some phthalimide containing analogues 2–12 have potential antimalarial activity. The compounds were evaluated against malaria strains 3D7 and K1, and cytotoxicity was measured against a mammalian cell line panel. A promising lead, compound 3, showed potent activity with an EC50 = 86 nM (3D7 strain), 201 nM (K1 strain) and low cytotoxicity in mammalian cells (SI > 290). Some structure–activity relationships have been identified for the antimalarial activity in these abietane analogues. 相似文献
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L. A. Fonina E. V. Kudryavtseva Zh. D. Bespalova M. A. Efremov A. A. Mikhailova E. A. Kirilina 《Russian Journal of Bioorganic Chemistry》2010,36(4):456-460
Bone marrow peptides Phe-Arg-Pro-Arg-Ile-Met-Thr-Pro (MP-4) and Val-Asp-Pro-Pro (MP-6) have been synthesized by the classical and solid-phase methods of peptide chemistry, and their differentiating activity has been studied on leukemia cell lines HL-60 and K-562. It has been shown that both peptides induce the terminal differentiation of leukemic blasts; however, their mechanisms of action are different. 相似文献
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A bacterial strain capable of utilizing E-pyridine-3-aldoxime as a nitrogen source was isolated from soil after a 4-month acclimation period and was identified as
Rhodococcus sp. The strain contained a novel aldoxime dehydration activity that catalyzed a stoichiometric dehydration of E-pyridine-3-aldoxime to form 3-cyanopyridine. The enzyme activity was induced by various aldoximes and nitriles. The strain
metabolized the aldoxime as follows: E-pyridine-3-aldoxime was dehydrated to form 3-cyanopyridine, which was converted to nicotinamide by a nitrile hydratase,
and the nicotinamide was successively hydrolyzed to nicotinic acid by an amidase.
Received: 21 January 1998 / Accepted: 12 May 1998 相似文献
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Francisco M. Garibotto Adriana D. Garro Marcelo F. Masman Ana M. Rodríguez Paul G.M. Luiten Marcela Raimondi Susana A. Zacchino Csaba Somlai Botond Penke Ricardo D. Enriz 《Bioorganic & medicinal chemistry》2010,18(1):158-167
The synthesis, in vitro evaluation, and conformational study of a new series of small-size peptides acting as antifungal agents are reported. In a first step of our study we performed a conformational analysis using Molecular Mechanics calculations. The electronic study was carried out using Molecular electrostatic potentials (MEPs) obtained from RHF/6-31G calculations. On the basis of the theoretical predictions three small-size peptides, RQWKKWWQWRR-NH2, RQIRRWWQWRR-NH2, and RQIRRWWQW-NH2 were synthesized and tested. These peptides displayed a significant antifungal activity against human pathogenic strains including Candida albicans and Cryptococcus neoformans. Our experimental and theoretical results allow the identification of a topographical template which can serve as a guide for the design of new compounds with antifungal properties for potential therapeutic applications against these pathogenic fungi. 相似文献
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Manju Sarkar Bimal Kumar Bachhawat Chitra Mandal 《Archives of biochemistry and biophysics》1984,233(1):286-289
An electrophoretically homogeneous agglutinin was purified from the albumin gland of Achatina fulica snails using asialofetunin-Sepharose 4B as an affinity column. The agglutinin was found to be temperature sensitive; it agglutinated rabbit and human umbilical cord erythrocytes only at low temperature. It was found to be specific for methyl-β-d-galactoside, and the best inhibitor was N-acetyllactosamine. 相似文献
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38 healthy subjects 22-28 years of age underwent the cold-pressor-test seven times on each of 21 consecutive days always at the same time of day. The subjective sensations of cold pain and the objective circulatory reactions were registered. There was observed only a very distinct subjective cold pain habituation from one exposition to the other. The habituation effect on blood pressure reaction however was of expected extent. The long term adaptation showed to have effect on the predominance of the two components of pain perception the epicritic and protopathic pain experience. Initially the cold stimulus produced an overwhelmingly protopathic sufferenig whilst at the end of the three weeks of adaptation there remained only a reduced and localized epicritic pain sensation. Typological differences of pain reactions and pain adaptation are discussed. 相似文献
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Daisuke Hinode Kaname Masuda Masami Yoshioka Kiyoko Watanabe Toshio Umemoto Daniel Grenier Denis Mayrand Ryo Nakamura 《FEMS microbiology letters》1995,131(2):211-217
Abstract A monoclonal antibody (mAb-PC) was produced against a BA p NA-hydrolyzing protease possessing hemagglutinating activity (Pase-C) from Porphyromonas gingivalis . Other P. gingivalis BA p NA-hydrolyzing enzymes (Pase-B and Pase-S) did not react with this antibody. By ELISA or SDS-PAGE and Western immunoblotting analysis, mAb-PC recognized all P. gingivalis and P. endodontalis strains tested but did not recognize other members of the Porphyromonas genus nor other putative periodontopathogenic organisms. Pase-C, extracellular vesicles (ECV) and human strains of P. gingivalis showed two major immunoreactive bands (44 kDa and 40 kDa), whereas a different pattern was obtained with animal strains of P. gingivalis . Biotinylarginyl chloromethane, an irreversible inhibitor of trypsin-like proteases, did not affect the reactivity of Pase-C with mAb-PC on immunoblot. By reversed-phase electronmicroscopy following immunogold labeling, the antibody was shown to bind to the cell surface of P. gingivalis . mAb-PC inhibited the hemagglutinating activity of both P. gingivalis cells and ECV whereas a monoclonal antibody against LPS of P. gingivalis did not. These results suggest that Pase-C is located on the cell surface of P. gingivalis and may participate in erythrocyte binding. 相似文献
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Burgeson JR Moore AL Boutilier JK Cerruti NR Gharaibeh DN Lovejoy CE Amberg SM Hruby DE Tyavanagimatt SR Allen RD Dai D 《Bioorganic & medicinal chemistry letters》2012,22(13):4263-4272
A series of acylthiourea derivatives were designed, synthesized, and evaluated for broad-spectrum antiviral activity with selected viruses from Poxviridae (vaccinia virus) and two different genera of the family Bunyaviridae (Rift Valley fever and La Crosse viruses). A compound selected from a library screen, compound 1, displayed submicromolar antiviral activity against both vaccinia virus (EC(50)=0.25 μM) and La Crosse virus (EC(50)=0.27 μM) in cytopathic effect (CPE) assays. SAR analysis was performed to further improve antiviral potency and to optimize drug-like properties of the initial hits. During our analysis, we identified 26, which was found to be nearly fourfold more potent than 1 against both vaccinia and La Crosse viruses. Selected compounds were further tested to more fully characterize the spectrum of antiviral activity. Many of these possessed single digit micromolar and sub-micromolar antiviral activity against a diverse array of targets, including influenza virus (Orthomyxoviridae), Tacaribe virus (Arenaviridae), and dengue virus (Flaviviridae). 相似文献
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Physicochemical studies of binding of 4-methylumbelliferyl beta-D-galactopyranoside to cold agglutinin.
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The fluorescence of 4-methylumbelliferyl beta-D-galactopyranoside (MeUmbGalp) was quenched in the presence of cold agglutinin, showing that there was binding between MeUmbGalp and cold agglutinin. That binding was saccharide-specific. By using this quenching phenomenon, the association constants (Ka) of the binding of cold agglutinin at different temperatures (10 degrees C and 15 degrees C) to MeUmbGalp and also the number of binding sites were calculated. The Ka values were found to be 2.63 x 10(3) M-1 at 10 degrees C and 1.58 x 10(3) M-1 at 15 degrees C. Though there is a change in Ka values, the number of binding sites was calculated to be six at both temperatures (10 degrees C and 15 degrees C). From the Ka values the thermodynamic parameters (free energy, enthalpy and entropy) of the binding were derived, and analysis of the data indicated that the binding is spontaneous, exothermic and hydrophobic in nature. 相似文献
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Li-Jun Ling Yan-Zhuo Yang Yu-Rong Bi 《World journal of microbiology & biotechnology》2010,26(3):545-554
Pinellia ternata agglutinin (PTA) from P. ternata f. angustata is a two-domain GNA-related lectin. The current study indicates that the PTA gene encodes a precursor consisting of two tandemly arrayed domains, N-terminal domain (PTA-DOM1) and C-terminal domain (PTA-DOM2). Both domains and the precursor without signal peptide (PTA-P) present different number of activity mannose-binding sites which play key roles for the lectin function. Analyses of the three fusion proteins, PTA-DOM1, PTA-DOM2 and PTA-P, expressed in Escherichia coli revealed that one mannose-binding site the agglutination activity while the additional sites do not possess such activity. However, the number of carbohydrate-binding sites suggests some significant properties on the antifungal effectiveness. In addition, each of the PTA domains has the same function when compared with the natural PTA (N-PTA). The information on PTA gene obtained in this study will served as baseline information in developing this protein as a form of transgenic plant protection. 相似文献
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The cold agglutinin from the albumin gland of the snail Achatina fulica was purified to homogeneity by using sheep gastric mucin-Sepharose 4B as affinity column followed by gel filtration on Bio-Gel P-300. The homogeneity was checked by alkaline gel electrophoresis, immunodiffusion and immunoelectrophoresis. The purified cold agglutinin is a glycoprotein of native M2 220,000 consisting of three non-covalently bound subunits of Mr 84,000, 74,000 and 62,000 and having a pI value of 4.5. The predominant amino acids are aspartic acid and glutamic acid (or amides) and serine, which account for 39% of the residues. About 3% of the residues are half-cystine. The lectin is a glycoprotein with about 30.7% carbohydrate, the most abundant sugars being galactose, N-acetylgalactosamine and N-acetylglucosamine. Mannose, xylose and fucose are also present. The inhibition of agglutination of human umbilical-cord erythrocytes by the cold agglutinin is specific for methyl beta-D-galactoside and also for glycolipids present on cord erythrocytes. The c.d. data show only negative ellipticity values in the far-u.v. region for the protein at various concentrations and temperatures and also in the presence of the hapten lactose (at different concentrations), indicating the presence of a random-coil conformation in the agglutinin that varies according to temperature. 相似文献
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Heikkinen J Risteli M Wang C Latvala J Rossi M Valtavaara M Myllylä R 《The Journal of biological chemistry》2000,275(46):36158-36163
Lysyl hydroxylase (EC ) and glucosyltransferase (EC ) are enzymes involved in post-translational modifications during collagen biosynthesis. We reveal in this paper that the protein produced by the cDNA for human lysyl hydroxylase isoform 3 (LH3) has both lysyl hydroxylase and glucosyltransferase (GGT) activities. The other known lysyl hydroxylase isoforms, LH1, LH2a, and LH2b, have no GGT activity. Furthermore, antibodies recognizing the amino acid sequence of human LH3 and those against a highly purified chicken GGT partially inhibited the GGT activity. Similarly, a partial inhibition was observed when these antibodies were tested against GGT extracted from human skin fibroblasts. In vitro mutagenesis experiments demonstrate that the amino acids involved in the GGT active site differ from those required for LH3 activity. 相似文献