Endohexosaminidase-catalysed glycosylation with oxazoline donors: Fine tuning of catalytic efficiency and reversibility

Thomas W. D. E. Rising; Christoph D. Heidecke; James W. B. Moir; Zhenlian Ling; Antony J. Fairbanks

doi:10.1002/chem.200800365

Endohexosaminidase-catalysed glycosylation with oxazoline donors: Fine tuning of catalytic efficiency and reversibility

Thomas W. D. E. Rising, Christoph D. Heidecke, James W. B. Moir, Zhenlian Ling, Antony J. Fairbanks

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

A complete series of oxazoline di-, tri-, tetra-, and hexasaccharides, corresponding to the core sections of N-linked glycoprotein high mannose glycans, together with the corresponding oligosaccharides containing a central glucose unit, were synthesised and tested as glycosyl donors for glycosylation of a GlcNAcAsn glycosyl amino acid catalysed by the endohexosaminidases M (Endo M), A (Endo A) and H (Endo H). Whilst Endo H did not catalyse any glycosylation reactions, both Endo M and Endo A efficiently catalysed glycosylations that were not limited to donors containing the Man beta(1 -> 4)GlcNAc linkage. Precise structure activity relationships and time course studies have revealed fine-tuning of the efficiency of the synthetic processes which correlated both with the enzyme used and the precise oxazoline structure. Efficient irreversible glycosylation was achievable with both Endo M and Endo A, further demonstrating the use of structurally modified oxazoline donors as transition state mimics in order to promote enzyme-catalysed synthesis, whilst precluding product hydrolysis; enzymes in these cases display "glycoligase" activity.

Original language	English
Pages (from-to)	6444-6464
Number of pages	21
Journal	Chemistry - A European Journal
Volume	14
Issue number	21
DOIs	https://doi.org/10.1002/chem.200800365
Publication status	Published - 2008

Keywords

carbohydrates
enzyme catalysis
glycopeptides
glycoproteins
glycosylation
BETA-N-ACETYLGLUCOSAMINIDASE
CHEMOENZYMATIC SYNTHESIS
GLYCOPROTEIN-SYNTHESIS
OLIGOSACCHARIDE-TRANSFER
GLYCOPEPTIDES
SUBSTRATE
TRANSGLYCOSYLATION
GLYCOFORMS
MOIETIES
GLYCANS

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10.1002/chem.200800365

Cite this

@article{c07a49bfa37c4761bf385bc06da867db,

title = "Endohexosaminidase-catalysed glycosylation with oxazoline donors: Fine tuning of catalytic efficiency and reversibility",

abstract = "A complete series of oxazoline di-, tri-, tetra-, and hexasaccharides, corresponding to the core sections of N-linked glycoprotein high mannose glycans, together with the corresponding oligosaccharides containing a central glucose unit, were synthesised and tested as glycosyl donors for glycosylation of a GlcNAcAsn glycosyl amino acid catalysed by the endohexosaminidases M (Endo M), A (Endo A) and H (Endo H). Whilst Endo H did not catalyse any glycosylation reactions, both Endo M and Endo A efficiently catalysed glycosylations that were not limited to donors containing the Man beta(1 -> 4)GlcNAc linkage. Precise structure activity relationships and time course studies have revealed fine-tuning of the efficiency of the synthetic processes which correlated both with the enzyme used and the precise oxazoline structure. Efficient irreversible glycosylation was achievable with both Endo M and Endo A, further demonstrating the use of structurally modified oxazoline donors as transition state mimics in order to promote enzyme-catalysed synthesis, whilst precluding product hydrolysis; enzymes in these cases display {"}glycoligase{"} activity.",

keywords = "carbohydrates, enzyme catalysis, glycopeptides, glycoproteins, glycosylation, BETA-N-ACETYLGLUCOSAMINIDASE, CHEMOENZYMATIC SYNTHESIS, GLYCOPROTEIN-SYNTHESIS, OLIGOSACCHARIDE-TRANSFER, GLYCOPEPTIDES, SUBSTRATE, TRANSGLYCOSYLATION, GLYCOFORMS, MOIETIES, GLYCANS",

author = "Rising, {Thomas W. D. E.} and Heidecke, {Christoph D.} and Moir, {James W. B.} and Zhenlian Ling and Fairbanks, {Antony J.}",

year = "2008",

doi = "10.1002/chem.200800365",

language = "English",

volume = "14",

pages = "6444--6464",

journal = "Chemistry - A European Journal",

issn = "0947-6539",

publisher = "Wiley-Blackwell",

number = "21",

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TY - JOUR

T1 - Endohexosaminidase-catalysed glycosylation with oxazoline donors: Fine tuning of catalytic efficiency and reversibility

AU - Rising, Thomas W. D. E.

AU - Heidecke, Christoph D.

AU - Moir, James W. B.

AU - Ling, Zhenlian

AU - Fairbanks, Antony J.

PY - 2008

Y1 - 2008

N2 - A complete series of oxazoline di-, tri-, tetra-, and hexasaccharides, corresponding to the core sections of N-linked glycoprotein high mannose glycans, together with the corresponding oligosaccharides containing a central glucose unit, were synthesised and tested as glycosyl donors for glycosylation of a GlcNAcAsn glycosyl amino acid catalysed by the endohexosaminidases M (Endo M), A (Endo A) and H (Endo H). Whilst Endo H did not catalyse any glycosylation reactions, both Endo M and Endo A efficiently catalysed glycosylations that were not limited to donors containing the Man beta(1 -> 4)GlcNAc linkage. Precise structure activity relationships and time course studies have revealed fine-tuning of the efficiency of the synthetic processes which correlated both with the enzyme used and the precise oxazoline structure. Efficient irreversible glycosylation was achievable with both Endo M and Endo A, further demonstrating the use of structurally modified oxazoline donors as transition state mimics in order to promote enzyme-catalysed synthesis, whilst precluding product hydrolysis; enzymes in these cases display "glycoligase" activity.

AB - A complete series of oxazoline di-, tri-, tetra-, and hexasaccharides, corresponding to the core sections of N-linked glycoprotein high mannose glycans, together with the corresponding oligosaccharides containing a central glucose unit, were synthesised and tested as glycosyl donors for glycosylation of a GlcNAcAsn glycosyl amino acid catalysed by the endohexosaminidases M (Endo M), A (Endo A) and H (Endo H). Whilst Endo H did not catalyse any glycosylation reactions, both Endo M and Endo A efficiently catalysed glycosylations that were not limited to donors containing the Man beta(1 -> 4)GlcNAc linkage. Precise structure activity relationships and time course studies have revealed fine-tuning of the efficiency of the synthetic processes which correlated both with the enzyme used and the precise oxazoline structure. Efficient irreversible glycosylation was achievable with both Endo M and Endo A, further demonstrating the use of structurally modified oxazoline donors as transition state mimics in order to promote enzyme-catalysed synthesis, whilst precluding product hydrolysis; enzymes in these cases display "glycoligase" activity.

KW - carbohydrates

KW - enzyme catalysis

KW - glycopeptides

KW - glycoproteins

KW - glycosylation

KW - BETA-N-ACETYLGLUCOSAMINIDASE

KW - CHEMOENZYMATIC SYNTHESIS

KW - GLYCOPROTEIN-SYNTHESIS

KW - OLIGOSACCHARIDE-TRANSFER

KW - GLYCOPEPTIDES

KW - SUBSTRATE

KW - TRANSGLYCOSYLATION

KW - GLYCOFORMS

KW - MOIETIES

KW - GLYCANS

U2 - 10.1002/chem.200800365

DO - 10.1002/chem.200800365

M3 - Article

SN - 0947-6539

VL - 14

SP - 6444

EP - 6464

JO - Chemistry - A European Journal

JF - Chemistry - A European Journal

IS - 21

ER -