In Vitro activity of Neisseria meningitidis PglL O- oligosaccharyltransferase with diverse synthetic lipid donors and a UDP-activated sugar

Matias A. Musumeci, Isabelle Hug, Nichollas E. Scott, M. Veronica Ielmini, Leonard J. Foster, Peng G. Wang, Mario F. Feldman

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Oligosaccharyltransferases (OTases) are enzymes that catalyze the transfer of an oligosaccharide from a lipid carrier to an acceptor molecule, commonly a protein. OTases are classified as N-OTases and O-OTases, depending on the nature of the glycosylation reaction. The N-OTases catalyze the glycan transfer to amide groups in asparagines in a reaction named N-linked glycosylation. The O-OTases are responsible for protein O-linked glycosylation, which involves the attachment of glycans to hydroxyl groups of serine or threonine residues. These enzymes exhibit a relaxed specificity and are able to transfer a variety of glycan structures to different protein acceptors. This property confers OTases with great biotechnological potential as these enzymes can produce glycoconjugates relevant to the pharmaceutical industry. Furthermore, OTases are thought to be involved in pathogenesis mechanisms. Several aspects of the functionality of OTases are not fully understood. In this work, we developed a novel approach to perform kinetic studies on PglL, theO-OTase from Neisseria meningitidis.Weinvestigated the importance of the acyl moiety of the lipid glycan donor substrate on the functionality of PglL by testing the efficiency of glycosylation reactions using synthetic substrates carrying the same glycan structure but different acyl moieties.Wefound that PglL can function with many lipids as glycan donors, although the length and the conformation of the lipid moiety significantly influenced the catalytic efficiency. Interestingly, PglL was also able to transfer a monosaccharide employing its nucleotide-activated form, acting as a Leloir glycosyltransferase. These results provide new insights on the function and the evolution of oligosaccharyltransferases.

Original languageEnglish
Pages (from-to)10578-10587
Number of pages10
JournalJournal of Biological Chemistry
Volume288
Issue number15
DOIs
StatePublished - Apr 12 2013
Externally publishedYes

Fingerprint Dive into the research topics of 'In Vitro activity of Neisseria meningitidis PglL O- oligosaccharyltransferase with diverse synthetic lipid donors and a UDP-activated sugar'. Together they form a unique fingerprint.

  • Cite this