共查询到20条相似文献,搜索用时 15 毫秒
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Kanae Sano Taiki Kuribara Nozomi Ishii Ayumi Kuroiwa Toshitada Yoshihara Seiji Tobita Kiichiro Totani Ichiro Matsuo 《化学:亚洲杂志》2019,14(11):1965-1969
Golgi endo‐α‐mannosidase (G‐EM) catalyzes an alternative deglucosylation process for N‐glycans and plays important roles in the post‐endoplasmic reticulum (ER) quality control pathway. To understand the post‐ER quality control mechanism, we synthesized a tetrasaccharide probe for the detection of the hydrolytic activity of G‐EM based on a fluorescence quenching assay. The probe was labeled with an N‐methylanthraniloyl group as a reporter dye at the non‐reducing end and a 2,4‐dinitrophenyl group as a quencher at the reducing end. This probe is hydrolyzed to disaccharide derivatives by G‐EM, resulting in increased fluorescence intensity. Thus, the fluorescence signal is directly proportional to the amount of disaccharide derivative present, allowing the G‐EM activity to be evaluated easily and quantitatively. 相似文献
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Dr. Nathalie Lunau Dipl.‐Chem. Katrin Seelhorst Dipl.‐Chem. Stefanie Kahl Dr. Kathrin Tscherch Dr. Christina Stacke Prof. Dr. Sascha Rohn Prof. Dr. Joachim Thiem Prof. Dr. Ulrich Hahn Prof. Dr. Chris Meier 《Chemistry (Weinheim an der Bergstrasse, Germany)》2013,19(51):17379-17390
Fucosylation is often the final process in glycan biosynthesis. The resulting glycans are involved in a variety of biological processes, such as cell adhesion, inflammation, or tumor metastasis. Fucosyltransferases catalyze the transfer of fucose residues from the activated donor molecule GDP‐β‐L ‐fucose to various acceptor molecules. However, detailed information about the reaction processes is still lacking for most fucosyltransferases. In this work we have monitored α1,3‐fucosyltransferase activity. For both donor and acceptor substrates, the introduction of a fluorescent ATTO dye was the last step in the synthesis. The subsequent conversion of these substrates into fluorescently labeled products by α1,3‐fucosyltransferases was examined by high‐performance thin‐layer chromatography coupled with mass spectrometry as well as dual‐color fluorescence cross‐correlation spectroscopy, which revealed that both fluorescently labeled donor GDP‐β‐L ‐fucose‐ATTO 550 and acceptor N‐acetyllactosamine‐ATTO 647N were accepted by recombinant human fucosyltransferase IX and Helicobacter pylori α1,3‐fucosyltransferase, respectively. Analysis by fluorescence cross‐correlation spectroscopy allowed a quick and versatile estimation of the progress of the enzymatic reaction and therefore this method can be used as an alternative method for investigating fucosyltransferase reactions. 相似文献
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Cover Picture: Synthesis of a Pentasaccharide and Neoglycoconjugates Related to Fungal α‐(1→3)‐Glucan and Their Use in the Generation of Antibodies to Trace Aspergillus fumigatus Cell Wall (Chem. Eur. J. 3/2015) 下载免费PDF全文
Dr. Bozhena S. Komarova Maria V. Orekhova Dr. Yury E. Tsvetkov Remi Beau Dr. Vishukumar Aimanianda Prof. Jean‐Paul Latgé Prof. Nikolay E. Nifantiev 《Chemistry (Weinheim an der Bergstrasse, Germany)》2015,21(3):917-917
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《Chemistry (Weinheim an der Bergstrasse, Germany)》2003,9(15):3453-3453
The cover picture shows a 96‐well plate illuminated by a UV lamp and viewed through a cutoff filter. All wells contain a Cy3.5‐BHQ1 (reporter quencher) dual‐labeled oligonucleotide probe in buffer. Complementary sequence was added to three columns. The probe is virtually nonfluorescent due to static quenching through formation of an intramolecular dimer. Addition of complement separates the dye and quencher, causing fluorescence. Intramolecular dimers as a new design strategy for fluorescence‐quenched probes is discussed by M. K. Johansson and R. M. Cook in the Concepts article on p. 3466 ff. 相似文献
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Toshiki Nokami Dr. Akito Shibuya Shino Manabe Dr. Yukishige Ito Dr. Jun‐ichi Yoshida Prof. Dr. 《Chemistry (Weinheim an der Bergstrasse, Germany)》2009,15(10):2252-2255
Low‐temperature electrochemical oxidation of thioglycosides gave glycosyl triflates from which glycosyl sulfonium ions were produced (see scheme). The latter were characterized by NMR spectroscopy and cold‐spray mass spectrometry as a mixture of α‐ and β‐isomers (45:55). The α‐glycosyl sulfonium ion exhibited higher reactivity than the β‐glycosyl sulfonium ion in the reaction with methanol, which gave a mixture of α‐ and β‐methyl glycosides (41:59).
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Matthew G. Alteen Christina Gros Richard W. Meek David A. Cardoso Jil A. Busmann Gontran Sangouard Matthew C. Deen Hong‐Yee Tan David L. Shen Cecilia C. Russell Gideon J. Davies Phillip J. Robinson Adam McCluskey David J. Vocadlo 《Angewandte Chemie (International ed. in English)》2020,59(24):9601-9609
Glycosyltransferases carry out important cellular functions in species ranging from bacteria to humans. Despite their essential roles in biology, simple and robust activity assays that can be easily applied to high‐throughput screening for inhibitors of these enzymes have been challenging to develop. Herein, we report a bead‐based strategy to measure the group‐transfer activity of glycosyltransferases sensitively using simple fluorescence measurements, without the need for coupled enzymes or secondary reactions. We validate the performance and accuracy of the assay using O‐GlcNAc transferase (OGT) as a model system through detailed Michaelis–Menten kinetic analysis of various substrates and inhibitors. Optimization of this assay and application to high‐throughput screening enabled screening for inhibitors of OGT, leading to a novel inhibitory scaffold. We believe this assay will prove valuable not only for the study of OGT, but also more widely as a general approach for the screening of glycosyltransferases and other group‐transfer enzymes. 相似文献