An overview of sphingolipid metabolism: from synthesis to breakdown.
Elevated Golgi pH impairs terminal -glycosylation by inducing mislocalization of Golgi glycosyltransferases
Understanding the stepwise synthesis of glycolipids.
Given the diversity and micro-heterogeneous nature of complex glycans, as well as the significant roles they play in biological processes, libraries of structurally well-defined glycans are in urgent demand for studies in glycobiology and glycomedicine. Decades of effort has been devoted to developing chemical methodologies and enzymatic approaches to synthesize N-glycans, however, none were able to (cost-)efficiently generate large numbers of N-glycans. This is mainly due to the facts that: (1) the chemical methodologies developed so far are not cost-effective nor rapid enough to prepare numerous complex N-glycans. Generally, each methodology has been developed specifically for certain N-glycan structures, and is thus not suitable for efficient synthesis of other complex N-glycans. (2) The GTs applied in the enzymatic or chemoenzymatic synthesis approaches were mostly from eukaryotes,,, which are typically hard to access and exhibit narrow substrate specificity towards glycan acceptors. Most recently, several N- and N-glycans with multisialylated poly-N-acetyllactosamine extensions were successfully synthesized using a single bacterial α2,6-sialyltransferase (Pd2,6ST). This revealed the power of bacterial GTs in diversifying glycans. (3) A highly efficient and rapid N-glycan purification approach is lacking. So far the only reported approach is gel-filtration, which takes many hours to separate each target, and is not able to separate complex glycans with only one monosaccharide difference.
Craven, F; Silva, J; Segarra-Maset, M; Huang, K; Both, P; Gough, J; Flitsch, S.L.; and Webb, S.; “‘One-pot’ sequential enzymatic modification of synthetic glycolipids in vesicle membranes” Chem. Commun., 2018, Accepted manuscript 4th Jan 2018
Biosynthesis and degradation of mammalian glycosphingolipids.
In order to demonstrate the usefulness of the protected core 4 O -glycan a segment (Gly 34 -Gly 58 ) of emmprin (extracellular matrix metalloproteinase inducer), a cancer metastasis-related glycoprotein, was synthesized by the solid-phase method, utilizing the pentasaccharyl Thr ( 2 ) to introduce an O -glycan in place of the native N -glycan at Asn 44 .
Denome SA, Elf PK, Henderson TA, Nelson DE and Young KD (1999) E. coli mutants lacking all possible combinations of eight penicillin binding proteins: viability, characteristics, and implications for peptidoglycan synthesis. Journal of Bacteriology 181: 3981–3993.
Glycosyltransferases: Tools for Synthesis and Modification of Glycans
L. L., Y. L., C. M. and P. G. W. designed experiments, analyzed data and wrote the manuscript. L. L. performed enzymatic synthesis, purification and HPLC identification of glycans; Y. L., B. W., and Z. X. performed chemical synthesis and NMR analysis; C. M., J. Q. and N. W. performed MS analysis. All other authors expressed and purified enzymes.
We have successfully developed a Core Synthesis/Enzymatic Extension (CSEE) strategy for efficient synthesis of structurally defined N-glycans and a HPLC-based approach for rapid purification of these compounds. The combination of CSEE and HPLC purification allowed rapid access to 0.5–2 mg of 73 homogenous N-glycans in high purities (>98%), including 63 isomers (21 groups). These N-glycans are valuable materials for glycan analysis and bioactivity evaluation. In this work, oligosaccharyl thioether was used as a chemical glycosylation donor for the convergent installation of branched GlcNAc-terminated antennae, this general and efficient approach produced 8 N-glycan core structures with high stereoselectivity and excellent overall yields. This work also demonstrated that any GlcNAc terminated glycans can be enzymatically extended to 5 or more longer glycans (including LeX and SLeX) using several robust glycosyltransferases. Moreover, this work showed that complex N-glycans are best purified using HPLC utilizing a HILIC column. In summary, the CSEE strategy described here provides a practical approach for rapid production of structurally defined N-glycans, and has the potential to become a general approach to solve the complexity and diversity of glycomes. This may mark the beginning of “mass production” of glycomes.
Synthesis of biantennary LacNAc-linked O -glycan (core …
Efficient Chemoenzymatic Synthesis of an N-glycan …
Effects of tunicamycin on the biosynthesis of glycosaminoglycans by embryonic chick cornea.
GlycoWorks RapiFluor-MS N-Glycan Kit : Waters
The participation of lipid-linked oligosaccharide in synthesis of membrane glycoproteins.
Waters has developed the GlycoWorks RapiFluor-MS N-Glycan Kit
Organization of the synthesis of glycolipid oligosaccharides in the Golgi complex.
Industrial & Applied Science
Isolation of a cytosolic beta-glucosidase from calf liver and characterization of its active site with alkyl glucosides and basic glycosyl derivatives.
Essentials of Glycobiology - NCBI Bookshelf
Sardzik, Robert; Sharma, Ritu; Kaloo, Sara; Voglmeir, Josef; Crocker, Paul R.; Flitsch, Sabine L. “Chemoenzymatic synthesis of sialooligosaccharides on arrays for studies of cell surface adhesion” CHEMICAL COMMUNICATIONS. 47(19); 5425 – 5427. – 2011
Publications | The Sabine L. Flitsch Research Group
Voglmeir, Josef; Kaloo, Sara; Laurent, Nicolas; Meloni, Marco M.; Bohlmann, Lisa; Wilson, Iain B. H.; Flitsch, Sabine L. “Biochemical correlation of activity of the alpha-dystroglycan-modifying glycosyltransferase POMGnT1 with mutations in muscle-eye-brain disease” BIOCHEMICAL JOURNAL. 436; 447 – 455. – JUN 1 2011
Flitsch Research Group - The Manchester Institute of Biotechnology.
Sardzik, Robert; Green, Anthony P.; Laurent, Nicolas; Both, Peter; Fontana, Carolina; Voglmeir, Josef; Weissenborn, Martin J.; Haddoub, Rose; Grassi, Paola; Haslam, Stuart M.; Widmalm, Goran; Flitsch, Sabine L. “Chemoenzymatic Synthesis of O-Mannosylpeptides in Solution and on Solid Phase” JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. 134(10); 4521 – 4524. – MAR 14 2012
Stable Isotope Products | Cambridge Isotope Laboratories
Inhibition by nojirimycin and 1-deoxynojirimycin of microsomal glucosidases from calf liver acting on the glycoprotein oligosaccharides Glc1-3Man9GlcNAc2.
Congenital disorder of glycosylation - Wikipedia
Olkhov, Rouslan V.; Weissenborn, Martin J.; Flitsch, Sabine L.; Shaw, Andrew M. “Glycosylation Characterization of Human and Porcine Fibrinogen Proteins by Lectin-Binding Biophotonic Microarray Imaging” ANALYTICAL CHEMISTRY. 86(1); 621 – 628. – JAN 7 2014
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