Enzymatic engineering of polysialic acid on cells in vitro and in vivo using a purified bacterial polysialyltransferase

Abderrahman El Maarouf, Damali Moyo Lee Yaw, Theresa Lindhout, Damien D Pearse, Warren Wakarchuk, Urs Rutishauser

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

In vertebrates, polysialic acid (PSA) is typically added to the neural cell adhesion molecule (NCAM) in the Golgi by PST or STX polysialyltransferase. PSA promotes plasticity, and its enhanced expression by viral delivery of the PST or STX gene has been shown to promote cellular processes that are useful for repair of the injured adult nervous system. Here we demonstrate a new strategy for PSA induction on cells involving addition of a purified polysialyltransferase from Neisseria meningitidis (PSTNm) to the extracellular environment. In the presence of its donor substrate (CMP-Neu5Ac), PSTNm synthesized PSA directly on surfaces of various cell types in culture, including Chinese hamster ovary cells, chicken DF1 fibroblasts, primary rat Schwann cells, and mouse embryonic stem cells. Similarly, injection of PSTNm and donor in vivo was able to produce PSA in different adult brain regions, including the cerebral cortex, striatum, and spinal cord. PSA synthesis by PSTNm requires the presence of the donor CMP-Neu5Ac, and the product could be degraded by the PSA-specific endoneuraminidase-N. Although PSTNm was able to add PSA to NCAM, most of its product was attached to other cell surface proteins. Nevertheless, the PSTNm-induced PSA displayed the ability to attenuate cell adhesion, promote neurite outgrowth, and enhance cell migration as has been reported for endogenous PSA-NCAM. Polysialylation by PSTNm occurred in vivo in less than 2.5 h, persisted in tissues, and then decreased within a few weeks. Together these characteristics suggest that a PSTNm-based approach may provide a valuable alternative to PST gene therapy.

Original languageEnglish
Pages (from-to)32770-32779
Number of pages10
JournalJournal of Biological Chemistry
Volume287
Issue number39
DOIs
StatePublished - Sep 21 2012

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Neisseria meningitidis
Neural Cell Adhesion Molecules
Cells
polysialic acid
In Vitro Techniques
Gene therapy
Schwann Cells
Cell adhesion
Neurology
Fibroblasts
Cricetulus
Stem cells
Cell culture
Cell Adhesion
Genetic Therapy
Cerebral Cortex
Nervous System
Cell Movement
Plasticity
Vertebrates

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Enzymatic engineering of polysialic acid on cells in vitro and in vivo using a purified bacterial polysialyltransferase. / El Maarouf, Abderrahman; Yaw, Damali Moyo Lee; Lindhout, Theresa; Pearse, Damien D; Wakarchuk, Warren; Rutishauser, Urs.

In: Journal of Biological Chemistry, Vol. 287, No. 39, 21.09.2012, p. 32770-32779.

Research output: Contribution to journalArticle

El Maarouf, Abderrahman ; Yaw, Damali Moyo Lee ; Lindhout, Theresa ; Pearse, Damien D ; Wakarchuk, Warren ; Rutishauser, Urs. / Enzymatic engineering of polysialic acid on cells in vitro and in vivo using a purified bacterial polysialyltransferase. In: Journal of Biological Chemistry. 2012 ; Vol. 287, No. 39. pp. 32770-32779.
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