Synthesis of homogeneous MUC1 oligomers via a bi-directional ligation strategy

Sheikha, Dima Al; Wilkinson, Brendan L.; Santhakumar, Gajan; Thaysen‐Andersen, Morten; Payne, Richard J.

doi:10.1039/c3ob41363b

Cited by 8 publications

(3 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It is a highly glycosylated transmembrane protein with an extracellular domain, which is, to a large extent, made up of a 20 amino acid sequence (referred to here as [1-20]MUC1: PPAHGVT-SAPDTRPAPGSTA) repeated in tandem 30 to 150 times. 11 This is an ideal model to validate new chemical methodologies 4,12 and a relevant biological target in tumor vaccine development. 13 Oxime-containing compounds are particularly well suited for building original immunoconjugates, the oxime bond being sufficiently hydrolytically-stable 14 to induce significant immune responses.…”

Section: Resultsmentioning

confidence: 99%

Solid phase oxime ligations for the iterative synthesis of polypeptide conjugates

et al. 2014

View full text Add to dashboard Cite

Peptide-based complex biomacromolecules are now optimally assembled by sequential ligation of unprotected peptide segments. However, this approach is still limited by the laborious chromatographic purification and handling steps needed for multiple successive chemoselective couplings, which leads to loss of material. An efficient alternative is solid phase chemical ligation (SPCL) initially developed for native chemical ligation. We report here an extension of this approach to iterative oxime ligation reactions, and describe a streamlined approach for the modular preparation of oxime-containing polypeptides. In particular, we determined optimal conditions to remove the Aloc group in the presence of aminooxy and oxime ether groups, and we extended the applicability of iterative C-to-N SPCL through simplification of the access to a C-terminally-grafted, unprotected peptide segment, using solid supported chemical transformations only. The high purity of the crude oxime-containing polypeptides highlights the efficiency of our approach.

show abstract

Section: Resultsmentioning

confidence: 99%

Solid phase oxime ligations for the iterative synthesis of polypeptide conjugates

et al. 2014

View full text Add to dashboard Cite

show abstract

“…A characteristic feature of mucins is that a unit sequence with multiple O ‐glycosylation sites is repeated many times . To synthesize a tandem repeat model of mucin, the SPPS of the repeat sequence, followed by the peptide ligation using the thioester method is an efficient strategy . The unit sequence of MUC1 mucin is STAPPAHGVTS*APDTRPAPG, in which we introduced sialyl Tn antigen at the S* residue.…”

Section: Figurementioning

confidence: 99%

Sialyl Tn Unit with TFA‐Labile Protection Realizes Efficient Synthesis of Sialyl Glycoprotein

Takeda

Takei

Asahina

et al. 2018

Chemistry A European J

View full text Add to dashboard Cite

Amino acids bearing 4-methylbenzyl (MBn) and 4-methoxybenzyl (MPM)-protected sialic acid were synthesized and used for the 9-fluorenylmethoxycarbonyl (Fmoc) solid-phase synthesis of a glycopeptide. The α-sialyl linkage of the MBn-protected unit was partially cleaved under the final deprotection by trifluoroacetic acid (TFA). In addition, the removal of several MBn groups were incomplete. On the other hand, the MPM-protected unit gave the desired glycopeptide without decomposition of the α-sialyl linkage. Using this unit, peptide thioesters of the tandem repeat unit of MUC1 mucin were synthesized by the N-alkylcysteine (NAC) method and used for the one-pot ligation by the thioester method. As a result, the three tandem repeats of MUC1 carrying sialyl Tn antigens were successfully synthesized.

show abstract

“…5,[49][50][51][52] Previous approaches to improve the understanding of MUC1 membrane trafficking and its interaction with other proteins, that both depend on its glycosylation and phosphorylation pattern, included the generation of mucin-type O-linked glycopeptides and proteins, employing chemical synthesis, as well as unnatural amino acid mutagenesis. 19,[53][54][55][56][57][58] However, these approaches have so far been limited by the size of the accessible proteins, the number of PTMs that can be introduced in a single construct and/or by the complexity of the glycosylation. Our chemoenzymatic method allows the synthesis of homogeneous MUC1 peptides that are site-specically O-glycosylated and can be linked to each other via chemoselective ligation reactions, generating longer, site-specically modied MUC1 sequences.…”

Section: Introductionmentioning

confidence: 99%

A quantitative and site-specific chemoenzymatic glycosylation approach for PEGylated MUC1 peptides

et al. 2014

View full text Add to dashboard Cite

Full control over complex post-translational modifications (PTMs), such as O-glycosylation, is a prerequisite for testing and understanding the biological role of these modifications in protein function. Despite considerable progress over the last years, high throughput and easy-to-use methods for the synthesis of complex glycosylated peptides are still missing. We present here an efficient methodology to produce homogeneous site-specifically O-glycosylated peptides. Sequential chemoenzymatic glycosylation and separation from the reaction components are achieved via the temporary attachment of a monodisperse polyethylene glycol (PEG) polymer to the N-terminus of these peptides. Subsequent proteolytic removal of the PEG moiety allows quantitative recovery of homogeneous O-glycopeptides, suitable as building blocks for glycoprotein synthesis. Here, we demonstrate the preparation of glucuronylated variants of MUC1, a well-known member of the human mucin family. Homogeneously O-glycosylated variants were synthesized and will be used to study the role of O-linked glucuronic acid epitopes within the functional environment of the human MUC1 tandem repeat.

show abstract

Synthesis of homogeneous MUC1 oligomers via a bi-directional ligation strategy

Cited by 8 publications

References 50 publications

Solid phase oxime ligations for the iterative synthesis of polypeptide conjugates

Solid phase oxime ligations for the iterative synthesis of polypeptide conjugates

Sialyl Tn Unit with TFA‐Labile Protection Realizes Efficient Synthesis of Sialyl Glycoprotein

A quantitative and site-specific chemoenzymatic glycosylation approach for PEGylated MUC1 peptides

Contact Info

Product

Resources

About