Glycopeptide and glycoprotein synthesis involving unprotected carbohydrate building blocks

Guo, Zhongwu; Shao, Ning

doi:10.1002/med.20033

Cited by 42 publications

(18 citation statements)

References 90 publications

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“…[8][9][10][11][12][13][14][15][16][17][18][19][20] In general, three major strategies have been applied for synthesizing homogeneous glycopeptides. The most common strategy is to incorporate pre-formed glycosyl amino acids as building blocks in conventional solid-phase or solution-phase peptide synthesis.…”

Section: Introductionmentioning

confidence: 99%

Chemoenzymatic synthesis of glycopeptides and glycoproteins through endoglycosidase-catalyzed transglycosylation

Wang¹

2008

Carbohydrate Research

124

113

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Homogeneous glycopeptides and glycoproteins are indispensable for detailed structural and functional studies of glycoproteins. It is also fundamentally important to have to correct glycosylation patterns for developing effective glycoprotein-based therapeutics. This review discusses a useful chemoenzymatic method that takes advantage of the endoglycosidase-catalyzed transglycosylation to attach an intact oligosaccharide to a polypeptide in a single step, without the need for any protecting groups. The exploration of sugar oxazolines (enzymatic reaction intermediates) as donor substrates has not only expanded substrate availability, but also has significantly enhanced the enzymatic transglycosylation efficiency. Moreover, the discovery of a novel mutant with glycosynthase-like activity has made it possible to synthesize homogeneous glycoproteins with full-size natural Nglycans. Recent advances in this highly convergent chemoenzymatic approach and its application for glycopeptide and glycoprotein synthesis are highlighted.

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Section: Introductionmentioning

confidence: 99%

Chemoenzymatic synthesis of glycopeptides and glycoproteins through endoglycosidase-catalyzed transglycosylation

Wang¹

2008

Carbohydrate Research

124

113

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“…The results obtained with ppGalNAcT1 for (MUC1) 2 could be extrapolated to a small protein of 172 aa composed of 8 MUC1 tandem repeats and a 6His tag, (MUC1) 8.…”

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confidence: 96%

“…The cycle can be easily scaled up and several mg of highly glycosylated (MUC1) 8 have been obtained which may be utilized for immunological studies in mice. The UDP-GalNAc regeneration cycle can probably be used by all GalNAc transferases with K m constants below 500 mm including EXT L2 and GM2 synthase.…”

mentioning

confidence: 99%

“…[6] Several approaches have been developed for their production, ranging from pure chemical to pure enzymatic methods. [7][8][9][10][11][12][13] Large-scale production using only chemical methods is difficult being both long and inefficient because of multiple protection/deprotection steps. Enzymatic synthesis of glycoconjugates is advantageous being fast and gives generally good yields.…”

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confidence: 99%

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Efficient Enzymatic Glycosylation of Peptides and Oligosaccharides from GalNAc and UTP

et al. 2006

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“…[2] We have already reported the synthesis of a glycoprotein with a homogeneous N-linked complex-type oligosaccharide.[3] This synthesis employed native chemical ligation (NCL) to perform peptide-segment coupling. NCL relies on the thiol-exchange reaction between a peptide with an a-thioester group at the C terminus and another peptide with a cysteine residue at the N terminus and on the subsequent intramolecular acyl transfer.…”

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confidence: 99%

Uncovering a Latent Ligation Site for Glycopeptide Synthesis

Okamoto

Kajihara

2008

Angew Chem Int Ed

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Glycosylation, one of the most important posttranslational modifications, plays an important role in a variety of biological events.[1] Oligosaccharides on glycoproteins exhibit structural heterogeneity, which makes it difficult to elucidate the relationship between the oligosaccharide structure and the function of the glycoprotein.Chemical synthesis is one of the powerful approaches for obtaining homogeneous glycoproteins. [2] We have already reported the synthesis of a glycoprotein with a homogeneous N-linked complex-type oligosaccharide.[3] This synthesis employed native chemical ligation (NCL) to perform peptide-segment coupling. NCL relies on the thiol-exchange reaction between a peptide with an a-thioester group at the C terminus and another peptide with a cysteine residue at the N terminus and on the subsequent intramolecular acyl transfer.[4] However, occasionally, the cysteine residue is not properly located or does not exist in the target protein. To take this potential difficulty into consideration, a long glycopeptide sequence that is 30-50 amino acids from one cysteine site to another cysteine site occasionally needs to be synthesized for glycoprotein synthesis by the NCL method. The synthesis of such a glycopeptide with an N-linked glycopeptide is not easy to perform [2,5] and requires an appropriate amount of N-linked complex-type oligosaccharides; therefore, there is greater difficulty in glycoprotein synthesis than in simple protein synthesis.To examine NCL without a cysteine residue in a long target peptide, reduction methods changing the sulfhydryl group of cysteine to a hydrogen atom after NCL and utilizing an auxiliary group have been developed. [6] In the latter method, the amino acid sequence at the ligation site is limited for performance. For the development of a widely usable method in glycopeptide synthesis, we have also explored suitable NCL approaches; this endeavor enabled us to find a new ligation position at the serine site in the consensus sequence NXS (X: any amino acid except for proline), by which an asparagine residue is generally incorporated in an Nlinked oligosaccharide. This sequence is found in glycoproteins along with the NXT sequence.[1] In order to use the serine site for a new NCL, we have examined the new concept and attempted the conversion of a cysteine residue into a serine residue after NCL. For such a technique, it was necessary to explore concise reaction sequences. As a result, we found the possibility of using a CNBr cleavage method at a methylcysteine site, which could be obtained by specific methylation of cysteine.[7] Herein, we report a new chemical ligation approach at serine sites, which relies on the conversion of a cysteine residue into a serine residue after NCL.The strategy is shown in Scheme 1. After NCL (product A), the conversion of cysteine into serine was performed by the following reactions: S methylation of cysteine with methyl 4-nitrobenzenesulfonate (product B) and intramolecular rearrangement by activation with CNBr in 80 % HCOOH solution...

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Glycopeptide and glycoprotein synthesis involving unprotected carbohydrate building blocks

Cited by 42 publications

References 90 publications

Chemoenzymatic synthesis of glycopeptides and glycoproteins through endoglycosidase-catalyzed transglycosylation

Chemoenzymatic synthesis of glycopeptides and glycoproteins through endoglycosidase-catalyzed transglycosylation

Efficient Enzymatic Glycosylation of Peptides and Oligosaccharides from GalNAc and UTP

Uncovering a Latent Ligation Site for Glycopeptide Synthesis

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