A recently described bangle lectin (PHL) from the bacterium Photorhabdus asymbiotica was identified as a mainly fucose-binding protein that could play an important role in the host-pathogen interaction and in the modulation of host immune response. Structural studies showed that PHL is a homo-dimer that contains up to seven L-fucose-specific binding sites per monomer. For these reasons, potential ligands of the PHL lectin: α-L-fucopyranosyl-containing mono-, di-, tetra-, hexa- and dodecavalent ligands were tested. Two types of polyvalent structures were investigated - calix[4]arenes and dendrimers. The shared feature of all these structures was a C-glycosidic bond instead of the more common but physiologically unstable O-glycosidic bond. The inhibition potential of the tested structures was assessed using different techniques - hemagglutination, surface plasmon resonance, isothermal titration calorimetry, and cell cross-linking. All the ligands proved to be better than free L-fucose. The most active hexavalent dendrimer exhibited affinity three orders of magnitude higher than that of standard L-fucose. To determine the binding mode of some ligands, crystal complex PHL/fucosides 2 - 4 were prepared and studied using X-ray crystallography. The electron density in complexes proved the presence of the compounds in 6 out of 7 fucose-binding sites.
- MeSH
- antibakteriální látky chemie farmakologie terapeutické užití MeSH
- bakteriální infekce farmakoterapie mikrobiologie MeSH
- bakteriální proteiny antagonisté a inhibitory chemie izolace a purifikace metabolismus MeSH
- dendrimery chemie farmakologie terapeutické užití MeSH
- erytrocyty MeSH
- fukosa analogy a deriváty farmakologie terapeutické užití MeSH
- hemaglutinace účinky léků MeSH
- interakce hostitele a patogenu účinky léků MeSH
- krystalografie rentgenová MeSH
- lektiny antagonisté a inhibitory chemie izolace a purifikace metabolismus MeSH
- lidé MeSH
- ligandy MeSH
- molekulární modely MeSH
- Photorhabdus metabolismus MeSH
- povrchová plasmonová rezonance MeSH
- rekombinantní proteiny chemie izolace a purifikace metabolismus MeSH
- vazebná místa MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Photorhabdus luminescens is known for its symbiosis with the entomopathogenic nematode Heterorhabditis bacteriophora and its pathogenicity toward insect larvae. A hypothetical protein from P. luminescens was identified, purified from the native source, and characterized as an l-fucose-binding lectin, named P. luminescens lectin (PLL). Glycan array and biochemical characterization data revealed PLL to be specific toward l-fucose and the disaccharide glycan 3,6-O-Me2-Glcβ1-4(2,3-O-Me2)Rhaα-O-(p-C6H4)-OCH2CH2NH2 PLL was discovered to be a homotetramer with an intersubunit disulfide bridge. The crystal structures of native and recombinant PLL revealed a seven-bladed β-propeller fold creating seven putative fucose-binding sites per monomer. The crystal structure of the recombinant PLL·l-fucose complex confirmed that at least three sites were fucose-binding. Moreover, the crystal structures indicated that some of the other sites are masked either by the tetrameric nature of the lectin or by incorporation of the C terminus of the lectin into one of these sites. PLL exhibited an ability to bind to insect hemocytes and the cuticular surface of a nematode, H. bacteriophora.
- MeSH
- amébová dyzenterie diagnóza etiologie komplikace MeSH
- amébový absces jater diagnóza etiologie komplikace MeSH
- cysteinové endopeptidasy izolace a purifikace škodlivé účinky MeSH
- entamébóza diagnóza etiologie komplikace MeSH
- Entamoeba histolytica izolace a purifikace patogenita růst a vývoj MeSH
- lektiny izolace a purifikace škodlivé účinky MeSH
- peptidy izolace a purifikace škodlivé účinky MeSH
- virulence fyziologie imunologie MeSH
- MeSH
- aglutinin z podzemnice olejné analýza MeSH
- biomedicínský výzkum metody MeSH
- histologické techniky metody využití MeSH
- indikátory a reagencie * MeSH
- lektiny analýza izolace a purifikace MeSH
- lidé MeSH
- monoklonální protilátky MeSH
- oligosacharidy analýza klasifikace MeSH
- proteiny analýza izolace a purifikace MeSH
- rostlinné lektiny analýza izolace a purifikace MeSH
- sacharidy * analýza klasifikace MeSH
- slezina * anatomie a histologie cytologie fyziologie MeSH
- statistika jako téma MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
The purple pigmented bacterium Chromobacterium violaceum is a dominant component of tropical soil microbiota that can cause rare but fatal septicaemia in humans. Its sequenced genome provides insight into the abundant potential of this organism for biotechnological and pharmaceutical applications and allowed an ORF encoding a protein that is 60% identical to the fucose binding lectin (PA-IIL) from Pseudomonas aeruginosa and the mannose binding lectin (RS-IIL) from Ralstonia solanacearum to be identified. The lectin, CV-IIL, has recently been purified from C. violaceum [Zinger-Yosovich, K., Sudakevitz, D., Imberty, A., Garber, N. C., and Gilboa-Garber, N. (2006) Microbiology 152, 457-463] and has been confirmed to be a tetramer with subunit size of 11.86 kDa and a binding preference for fucose. We describe here the cloning of CV-IIL and its expression as a recombinant protein. A complete structure-function characterization has been made in an effort to analyze the specificity and affinity of CV-IIL for fucose and mannose. Crystal structures of CV-IIL complexes with monosaccharides have yielded the molecular basis of the specificity. Each monomer contains two close calcium cations that mediate the binding of the monosaccharides, which occurs in different orientations for fucose and mannose. The thermodynamics of binding has been analyzed by titration microcalorimetry, giving dissociation constants of 1.7 and 19 microM for alpha-methyl fucoside and alpha-methyl mannoside, respectively. Further analysis demonstrated a strongly favorable entropy term that is unusual in carbohydrate binding. A comparison with both PA-IIL and RS-IIL, which have binding preferences for fucose and mannose, respectively, yielded insights into the monosaccharide specificity of this important class of soluble bacterial lectins.
- MeSH
- bakteriální proteiny genetika chemie izolace a purifikace MeSH
- Chromobacterium chemie metabolismus MeSH
- entropie MeSH
- financování organizované MeSH
- fukosa metabolismus MeSH
- krystalizace MeSH
- lektin vázající mannosu genetika chemie izolace a purifikace MeSH
- lektiny genetika chemie izolace a purifikace MeSH
- mannosa metabolismus MeSH
- molekulární modely MeSH
- rekombinantní proteiny metabolismus MeSH
- rozpustnost MeSH
- sekundární struktura proteinů MeSH
- senzitivita a specificita MeSH
- statická elektřina MeSH
- vápník chemie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- vodíková vazba MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- bezobratlí MeSH
- biologické markery MeSH
- biomedicínský výzkum MeSH
- epidermální buňky MeSH
- epidermis MeSH
- epitelové buňky cytologie klasifikace metabolismus MeSH
- financování organizované MeSH
- galektin 3 diagnostické užití izolace a purifikace klasifikace MeSH
- galektiny * diagnostické užití izolace a purifikace klasifikace MeSH
- hlasové řasy cytologie MeSH
- Langerhansovy buňky cytologie MeSH
- lektiny diagnostické užití izolace a purifikace klasifikace MeSH
- lidé MeSH
- ligandy MeSH
- rohovka cytologie MeSH
- rostliny MeSH
- skvamocelulární nádory genetika imunologie MeSH
- statistika jako téma MeSH
- ústní sliznice cytologie MeSH
- Check Tag
- lidé MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
Přeruš. str. : il., tab., grafy ; 32 cm
Serum lectins will be isolated and their concentrations in sera of healthy and immunodeficient individuals with recurrent infections will be quantitatively estimated. Aminoacid sequences of lectins binding mannan and/or phosphomannan will be compared.
Budou izolovány sérové lektiny a kvantitativně stanoveny jejich koncentrace v sérech zdravých a imunodeficitních jedinců s opakovanými infekcemi. Budou porovnány aminokyselinové sekvence lektinů vázajících manan a fosfomanan.
- MeSH
- baktericidní aktivita krve MeSH
- lektiny izolace a purifikace krev MeSH
- mannosafosfáty MeSH
- mannosidy MeSH
- vazba proteinů MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Konspekt
- Patologie. Klinická medicína
- NLK Obory
- alergologie a imunologie
- NLK Publikační typ
- závěrečné zprávy o řešení grantu IGA MZ ČR
