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Glycobiology Advance Access originally published online on December 15, 2006
Glycobiology 2007 17(3):249-260; doi:10.1093/glycob/cwl075
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Published by Oxford University Press 2006

Polysialic acid bioengineering of neuronal cells by N-acyl sialic acid precursor treatment

Robert A. Pon2, Nancy J. Biggs2 and Harold J. Jennings1,2

2 Institute for Biological Sciences, National Research Council of Canada, 100 Sussex Drive, Ottawa K1A 0R6, Canada

1 To whom correspondence should be addressed; Tel: +1 613 990 0821; Fax: +1 613 941 1327; e-mail: harry.jennings{at}nrc-cnrc.gc.ca

Received on February 24, 2006; revised on December 5, 2006; accepted on December 8, 2006

The inherent promiscuity of the polysialic acid (PSA) biosynthetic pathway has been exploited by the use of exogenous unnatural sialic acid precursor molecules to introduce unnatural modifications into cellular PSA, and has found applications in nervous system development and tumor vaccine studies. The sialic acid precursor molecules N-propionyl- and N-butanoyl-mannosamine (ManPr, ManBu) have been variably reported to affect PSA biosynthesis ranging from complete inhibition to de novo production of modified PSA, thus illustrating the need for further investigation into their effects. In this study, we have used a monoclonal antibody (mAb) 13D9, specific to both N-propionyl-PSA and N-butanoyl-PSA (NPrPSA and NBuPSA), together with flow cytometry, to study precursor-treated tumor cells and NT2 neurons at different stages of their maturation. We report that both ManPr and ManBu sialic acid precursors are metabolized and the resultant unnatural sialic acids are incorporated into de novo surface sialylglycoconjugates in murine and human tumor cells and, for the first time, in human NT2 neurons. Furthermore, neither precursor treatment deleteriously affected endogenous PSA expression; however, with NT2 cells, PSA levels were naturally downregulated as a function of their maturation into polarized neurons independent of sialic acid precursor treatment.

Key words: glycan bioengineering / human neurons / N-acyl mannosamines / polysialic acid / tumor cell


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