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91.
Eric Sterner Lingyun Li Priscilla Paul Julie M. Beaudet Jian Liu Robert J. Linhardt Jonathan S. Dordick 《Analytical and bioanalytical chemistry》2014,406(2):525-536
O-sulfotransferases (OSTs) are critical enzymes in the cellular biosynthesis of the biologically and pharmacologically important heparan sulfate and heparin. Recently, these enzymes have been cloned and expressed in bacteria for application in the chemoenzymatic synthesis of glycosaminoglycan-based drugs. OST activity assays have largely relied on the use of radioisotopic methods using [35S] 3′-phosphoadenosine-5′-phosphosulfate and scintillation counting. Herein, we examine alternative assays that are more compatible with a biomanufacturing environment. A high throughput microtiter-based approach is reported that relies on a coupled bienzymic colorimetric assay for heparan sulfate and heparin OSTs acting on polysaccharide substrates using arylsulfotransferase-IV and p-nitrophenylsulfate as a sacrificial sulfogroup donor. A second liquid chromatography-mass spectrometric assay, for heparan sulfate and heparin OSTs acting on structurally defined oligosaccharide substrates, is also reported that provides additional information on the number and positions of the transferred sulfo groups within the product. Together, these assays allow quantitative and mechanistic information to be obtained on OSTs that act on heparan sulfate and heparin precursors. Figure
Herapan sulfate O-sulfotranferase coupled enzyme colorimetric assay 相似文献
92.
Lee JK Chatzichristidi M Zakhidov AA Taylor PG DeFranco JA Hwang HS Fong HH Holmes AB Malliaras GG Ober CK 《Journal of the American Chemical Society》2008,130(35):11564-11565
An acid-sensitive semiperfluoroalkyl resorcinarene was synthesized, and its lithographic properties were evaluated. Its solubility in segregated hydrofluoroether solvents enables the patterning of delicate organic electronic materials. 相似文献
93.
Sipho E. Mavundla Gerald F. Malgas Priscilla Baker Emmanuel. I. Iwuoha 《Electroanalysis》2008,20(21):2347-2353
Novel nanophase hexagonal structured polyaniline (PANI) and poly(2,5‐dimethoxyanilines) (PDMA) were synthesized by oxidative polymerization involving the respective anilines and a mixture of ferric chloride and ammonium persulfate. The morphological, spectral and electrochemical characteristics of the polymers were determined from the results of SEM, FTIR, UV‐vis, TGA and cyclic voltammetry experiments. The hexagonal PANI and PDMA nanorods (15–200 nm diameter) exhibited very good thermal stabilities, losing only 10% of their weight on heating to 400 °C. Electrochemical data indicated a pernigraniline state of the polymers with formal potential, E°′, values of 394±6 mV and 400±1 mV, for PANI (conductance, C=0.37×10?3 S) and PDMA (conductance, C=2.02×10?3 S), respectively. The pernigraniline state was confirmed by sharp FTIR pernigraniline quinoidic peaks (PANI: 1414 cm?1; PDMA: 1157 cm?1), and UV‐vis absorption maxima at 340–370 nm (PANI) and 450–650 nm (PDMA) which are characteristic of charge transfer excitons of the quinoid structures of pernigraniline. 相似文献
94.
Lugo-Mas P Dey A Xu L Davin SD Benedict J Kaminsky W Hodgson KO Hedman B Solomon EI Kovacs JA 《Journal of the American Chemical Society》2006,128(34):11211-11221