Enzymes in the Synthesis of Glycoconjugates

Schmaltz, Ryan M.; Hanson, Sarah R.; Wong, Chi‐Huey

doi:10.1021/cr200113w

Cited by 249 publications

(203 citation statements)

References 612 publications

(940 reference statements)

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“…N-glycans play crucial roles in protein homeostasis, modulating protein stability (Culyba, et al, 2011; Hanson, et al, 2009; Hebert, et al, 2014; Imperiali and Rickert, 1995; Joao and Dwek, 1993; Price, et al, 2012; Wang, et al, 1996; Wormald and Dwek, 1999), chaperone-mediated folding (Hammond, et al, 1994; Hebert, et al, 2014; Helenius and Aebi, 2001; Jitsuhara, et al, 2002; Oliver, et al, 1997; Ou, et al, 1993; Schmaltz, et al, 2011; Ware, et al, 1995), vesicular trafficking (Lu, et al, 1997; Martina, et al, 1998), and endoplasmic reticulum-associated degradation (ERAD) (Parodi, 2000; Vembar and Brodsky, 2008). N-glycans also strongly influence biological function, including immune/inflammatory responses and circulating glycoprotein clearance, wherein carbohydrate binding proteins (lectins) recognize specific glycoforms of surface glycoproteins (Ahrens, 1993; Bertozzi and Kiessling, 2001; Chui, et al, 2001; Sorensen, et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity

Murray

Chen

Antonopoulos

et al. 2015

Chemistry & Biology

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SUMMARY N-glycosylation plays an important role in protein folding and function. Previous studies demonstrate that a phenylalanine residue introduced at the n-2 position relative to an Asn-Xxx-Thr/Ser N-glycosylation sequon increases the sequon’s glycan occupancy in insect cells. Here, we show that any aromatic residue at n-2 increases glycan occupancy in human cells, and we show that this effect is dependent upon oligosaccharyltransferase substrate preferences rather than differences in other cellular processing events such as degradation or trafficking. Moreover, aromatic residues at n-2 alter glycan processing in the Golgi, producing proteins with less complex N-glycan structures. These results demonstrate that manipulating the sequence space surrounding N-glycosylation sequons is useful both for controlling glycosylation efficiency, thus enhancing glycan occupancy, and for influencing the N-glycan structures produced.

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Section: Introductionmentioning

confidence: 99%

Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity

Murray

Chen

Antonopoulos

et al. 2015

Chemistry & Biology

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“…glycoproteins [36][37][38]. Two molecules of the same protein from the same tissue may have different glycans [39]. Changes in glycan identity in a glycoprotein can have a dramatic effect in pathological processes [4,17].…”

Section: Resultsmentioning

confidence: 99%

Electrochemical sensing of concanavalin A and ovalbumin interaction in solution

Vargová

Helma

Paleček

et al. 2016

Analytica Chimica Acta

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“…Given the difficulties to isolate homogeneous glycoforms from natural sources, synthesis of structurally well-defined, core-fucosylated glycopeptides and glycoproteins is essential for various glycomics studies aiming to further decipher the structural and functional impact of core-fucosylation 24–26 . In animals and humans, core-fucosylation is catalyzed solely by the mammalian α1,6-fucosyltransferase, FUT8 27,28 .…”

Section: Introductionmentioning

confidence: 99%

Designer α1,6-Fucosidase Mutants Enable Direct Core Fucosylation of Intact N-Glycopeptides and N-Glycoproteins

Zhu

Christopher

et al. 2017

J. Am. Chem. Soc.

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Core fucosylation of N-glycoproteins plays a crucial role in modulating the biological functions of glycoproteins. Yet, the synthesis of structurally well-defined, core-fucosylated glycoproteins remains a challenging task due to the complexity in multi-step chemical synthesis or the inability of the biosynthetic α1,6-fucosyltransferase (FUT8) to directly fucosylate full-size mature N-glycans in a chemoenzymatic approach. We report in this paper the design and generation of potential α1,6-fucosynthase and fucoligase for direct core-fucosylation of intact N-glycoproteins. We found that mutation at the nucleophilic residue (D200) did not provide a typical glycosynthase from this bacterial enzyme, but several mutants with mutation at the general acid/base residue E274 of the Lactobacillus casei α1,6-fucosidase, including E274A, E274S, and E274G, acted as efficient glycoligases that could fucosylate a wide variety of complex N-glycopeptides and intact glycoproteins by using α-fucosyl fluoride as a simple donor substrate. Studies on the substrate specificity revealed that the α1,6-fucosidase mutants could introduce an α1,6-fucose moiety specifically at the Asn-linked GlcNAc moiety not only to GlcNAc-peptide, but also to high-mannose and complex type N-glycans in the context of N-glycopeptides, N-glycoproteins, and intact antibodies. This discovery opens a new avenue to a wide variety of homogeneous, core-fucosylated N-glycopeptides and N-glycoproteins that are hitherto difficult to obtain for structural and functional studies.

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Enzymes in the Synthesis of Glycoconjugates

Cited by 249 publications

References 612 publications

Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity

Enhanced Aromatic Sequons Increase Oligosaccharyltransferase Glycosylation Efficiency and Glycan Homogeneity

Electrochemical sensing of concanavalin A and ovalbumin interaction in solution

Designer α1,6-Fucosidase Mutants Enable Direct Core Fucosylation of Intact N-Glycopeptides and N-Glycoproteins

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