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31.
A model system for evaluating macroporous supports containing immobilized whole cells in affinity Chromatographic applications
is described. Whole cells were immobilized in a polyacrylamide network in a two-step polymerization process. The affinity
system discussed consists of immobilized cells ofSaccharomyces cervisiae in the purification of lentil lectin fromLens culinaris. 相似文献
32.
为获得人MBL蛋白,并对其功能进行初步研究,用DNA重组法构建了组氨到标签融合原核表达质粒pET28(b)-MBL。将重组质粒转入大肠杆菌BL21(DE3),经IPTG在37℃条件下诱导培养,利用SDS-PAGE,Westem-blot检测目的蛋白的表达,用IMAC金属螯合层析柱对其进行纯化。成功地表达了重组MBL蛋白,纯化的MBL浓度约为844μg/mL,为制备MBL的基因工程抗体奠定了基础。 相似文献
33.
A highly sensitive bioassay based on silver-enhanced luminol electrogenerated chemiluminescence (ECL) is reported for the determination of concanavalin A lectin. A gold electrode modified with the mixed self-assembled monolayer of thiolated mannoside and mercaptohexanol was used to selectively capture a target lectin, concanavalin A, through the specific interaction between mannoside and concanavalin A. Mannoside-functionalized gold nanoparticles were further introduced to the opposite binding sites of the tetrameric concanavalin A to form a sandwich-type complex. Silver enhancement step was performed to coat the surface of mannose-stabilized gold nanoparticles with silver. The deposited silver was dissolved in an acidic solution and further neutralized. The resulting silver ions were finally detected with luminol electrogenerated chemiluminescence, in which the silver ions greatly enhanced the chemiluminescence intensity. The present electrogenerated chemiluminescence bioassay detected concanavalin A from 0.190 to 10.0?µg/mL (r2?=?0.999) with a detection limit of 0.146?µg/mL (signal to noise ratio?=?3), which is much lower compared to previously reported methods such as microgravimetry, surface plasmon resonance, and colorimetry. Furthermore, the present bioassay showed good selectivity over possible interfering lectin proteins. 相似文献
34.
Alexandra Muñoz‐Bonilla Orietta León Vanesa Bordegé Manuel Sánchez‐Chaves Marta Fernández‐García 《Journal of polymer science. Part A, Polymer chemistry》2013,51(6):1337-1347
The syntheses of a variety of amphiphilic block glycopolymers based on 2‐{[(D ‐glucosamin‐2‐N‐yl)carbonyl]oxy}ethyl acrylate and n‐butyl acrylate or methyl methacrylate by single‐electron transfer‐living radical polymerization (SET‐LRP) are described. In a first step, the homopolymerization of unprotected acrylic glycomonomer to obtain well‐controlled glycopolymers is studied. Posterior and based on these studies, di‐ and triblock glycopolymers were synthesized via SET‐LRP of the glycomonomer from different hydrophobic blocks, varying the hydrophilic block lengths. All the copolymers are characterized by nuclear magnetic resonance spectroscopy and GPC. Moreover, their water solution behavior by dynamic light scattering and their capacity of interaction with Concanavalin A lectin by turbidimetry are analyzed. The effect on the block glycopolymers behavior of hydrophobic block nature and the length of glycopolymer segments is evaluated. © 2012 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2013 相似文献
35.
Sushil Kumar Mishra Johan Sund Johan Åqvist Jaroslav Koča 《Journal of computational chemistry》2012,33(29):2340-2350
The linear interaction energy (LIE) method to compute binding free energies is applied to lectin‐monosaccharide complexes. Here, we calculate the binding free energies of monosaccharides to the Ralstonia solanacearum lectin (RSL) and the Pseudomonas aeruginosa lectin‐II (PA‐IIL). The standard LIE model performs very well for RSL, whereas the PA‐IIL system, where ligand binding involves two calcium ions, presents a major challenge. To overcome this, we explore a new variant of the LIE model, where ligand–metal ion interactions are scaled separately. This model also predicts the saccharide binding preference of PA‐IIL on mutation of the receptor, which may be useful for protein engineering of lectins. © 2012 Wiley Periodicals, Inc. 相似文献
36.
Akkaladevi Narahari Musti J. Swamy 《Journal of photochemistry and photobiology. B, Biology》2009,97(1):40-47
The exposure and accessibility of the tryptophan residues in the chitooligosaccharide-specific pumpkin (Cucurbita maxima) phloem exudate lectin (PPL) have been investigated by fluorescence spectroscopy. The emission λmax of native PPL, seen at 338 nm was red-shifted to 348 nm upon denaturation by 6 M Gdn.HCl in the presence of 10 mM β-mercaptoethanol, indicating near complete exposure of the tryptophan residues to the aqueous medium, whereas a blue-shift to 335 nm was observed in the presence of saturating concentrations of chitotriose, suggesting that ligand binding leads to a decrease in the solvent exposure of the tryptophan residues. The extent of quenching was maximum with the neutral molecule, acrylamide whereas the ionic species, iodide and Cs+ led to significantly lower quenching, which could be attributed to the presence of charged amino acid residues in close proximity to some of the tryptophan residues. The Stern–Volmer plot for acrylamide was linear for native PPL and upon ligand binding, but became upward curving upon denaturation, indicating that the quenching occurs via a combination of static and dynamic mechanisms. In time-resolved fluorescence experiments, the decay curves could be best fit to biexponential patterns, for native protein, in the presence of ligand and upon denaturation. In each case both lifetimes systematically decreased with increasing acrylamide concentrations, indicating that quenching occurs predominantly via a dynamic process. 相似文献
37.
Z. S. Khashimova N. N. Kuznetsova S. V. Levitskaya 《Chemistry of Natural Compounds》2000,36(3):318-320
Lectin was isolated from seeds of IndianDatura innoxia. Certain of its physicochemical properties were determined. Its hemaglutinizing activity was demonstrated. The effect of the obtained protein on proliferation of the hybrid KML cell line from murine B-16 melanoma was studied. Its mitogenic properties were demonstrated.Translated from Khimiya Prirodnykh Soedinenii, No. 3, pp. 257–258, May–June, 2000. 相似文献
38.
Switching of Bacterial Adhesion to a Glycosylated Surface by Reversible Reorientation of the Carbohydrate Ligand
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Insa Stamer Prof. Dr. Mikkel B. Thygesen Prof. Dr. Andreas Terfort Prof. Dr. Thisbe K. Lindhorst 《Angewandte Chemie (International ed. in English)》2014,53(52):14583-14586
The surface recognition in many biological systems is guided by the interaction of carbohydrate‐specific proteins (lectins) with carbohydrate epitopes (ligands) located within the unordered glycoconjugate layer (glycocalyx) of cells. Thus, for recognition, the respective ligand has to reorient for a successful matching event. Herein, we present for the first time a model system, in which only the orientation of the ligand is altered in a controlled manner without changing the recognition quality of the ligand itself. The key for this orientational control is the embedding into an interfacial system and the use of a photoswitchable mechanical joint, such as azobenzene. 相似文献
39.
Maria Pia Lenza Iker Oyenarte Dr. Tammo Diercks Jon Imanol Quintana Dr. Ana Gimeno Dr. Helena Coelho Ana Diniz Dr. Francesca Peccati Sandra Delgado Alexandre Bosch Dr. Mikel Valle Dr. Oscar Millet Prof. Dr. Nicola G. A. Abrescia Dr. Asís Palazón Dr. Filipa Marcelo Dr. Gonzalo Jiménez-Osés Prof. Dr. Jesús Jiménez-Barbero Dr. Ana Ardá Dr. June Ereño-Orbea 《Angewandte Chemie (Weinheim an der Bergstrasse, Germany)》2020,132(52):23971-23979
The glycan structures of the receptor binding domain of the SARS-CoV2 spike glycoprotein expressed in human HEK293F cells have been studied by using NMR. The different possible interacting epitopes have been deeply analysed and characterized, providing evidence of the presence of glycan structures not found in previous MS-based analyses. The interaction of the RBD 13C-labelled glycans with different human lectins, which are expressed in different organs and tissues that may be affected during the infection process, has also been evaluated by NMR. In particular, 15N-labelled galectins (galectins-3, -7 and -8 N-terminal), Siglecs (Siglec-8, Siglec-10), and C-type lectins (DC-SIGN, MGL) have been employed. Complementary experiments from the glycoprotein perspective or from the lectin's point of view have permitted to disentangle the specific interacting epitopes in each case. Based on these findings, 3D models of the interacting complexes have been proposed. 相似文献
40.
Dr. Marco Mende Alexandra Tsouka Jasmin Heidepriem Grigori Paris Dr. Daniela S. Mattes Dr. Stephan Eickelmann Dr. Vittorio Bordoni Dr. Robert Wawrzinek Felix F. Fuchsberger Prof. Dr. Peter H. Seeberger Dr. Christoph Rademacher Dr. Martina Delbianco Dr. Alvaro Mallagaray Dr. Felix F. Loeffler 《Chemistry (Weinheim an der Bergstrasse, Germany)》2020,26(44):9954-9963
Single glycan–protein interactions are often weak, such that glycan binding partners commonly utilize multiple, spatially defined binding sites to enhance binding avidity and specificity. Current array technologies usually neglect defined multivalent display. Laser-based array synthesis technology allows for flexible and rapid on-surface synthesis of different peptides. By combining this technique with click chemistry, neo-glycopeptides were produced directly on a functionalized glass slide in the microarray format. Density and spatial distribution of carbohydrates can be tuned, resulting in well-defined glycan structures for multivalent display. The two lectins concanavalin A and langerin were probed with different glycans on multivalent scaffolds, revealing strong spacing-, density-, and ligand-dependent binding. In addition, we could also measure the surface dissociation constant. This approach allows for a rapid generation, screening, and optimization of a multitude of multivalent scaffolds for glycan binding. 相似文献