A quantitative and site-specific chemoenzymatic glycosylation approach for PEGylated MUC1 peptides — ASN Events
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  2. Session 5 SP Protein Synthesis
  3. A quantitative and site-specific chemoenzymatic glycosylation approach for PEGylated MUC1 peptides

A quantitative and site-specific chemoenzymatic glycosylation approach for PEGylated MUC1 peptides (#18)

Christian F.W. Becker 1 , Claudia Bello 1 , Karine Farbiarz 1 , Tilo Schwientek 2
  1. University of Vienna, Vienna, Austria
  2. Medical Biochemistry II, University of Cologne, Cologne

Full control over complex posttranslational modifications (PTMs), such as O-glycosylation, is a prerequisite for testing and understanding the biological role of these modifications. Many methods have been developed to achieve the in vitro synthesis of complex glycosylated peptides and proteins, either by synthetic or by using enzyme-based approaches.1-3 Despite the tremendous progress in liquid as well as in solid phase synthesis of complex carbohydrates4 the now available methods are still not as easy-to-use as the automated synthesis of oligonucleotides or peptides.

We present here an efficient methodology to produce site-specifically O-glycosylated peptides. Sequential chemoenzymatic glycosylation and separation from reaction components is achieved via temporary attachment of a polyethylene glycol (PEG) polymer5-7. Subsequent proteolytic removal of the PEG moiety allows quantitative recovery of glycopeptides suitable as building blocks for glycoprotein synthesis via chemoselective ligations. Here we demonstrate the preparation of glucuronylated variants of MUC1, a well-known member of the human mucin family.

References

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3.         C. Unverzagt and Y. Kajihara, Chemi. Soc. Rev., 2013, 42, 4408-4420.

4.         S. Serna, J. Etxebarria, N. Ruiz, M. Martin-Lomas, N. C. Reichardt, Chemistry, 2010, 16, 13163-13175.

5.         E. Bayer and M. Mutter, Nature, 1972, 237, 512-513.

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7.         T. Matsushita, I. Nagashima, M. Fumoto, T. Ohta, K. Yamada, H. Shimizu, H. Hinou, K. Naruchi, T. Ito, H. Kondo, S. Nishimura, J. Am. Chem. Soc., 2010, 132, 16651-16656.