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

   


Authors: El-Maarouf, A. ; Yaw, D. M. L.; Lindhout, T.; Pearse, D. D.; Wakarchuk, W.; Rutishauser, U.
Article Title: Enzymatic engineering of polysialic acid on cells in vitro and in vivo using a purified bacterial polysialyltransferase
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 (PST Nm) to the extracellular environment. In the presence of its donor substrate (CMP-Neu5Ac), PST Nm 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 PST Nm 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 PST Nm requires the presence of the donor CMP-Neu5Ac, and the product could be degraded by the PSA-specific endoneuraminidase-N. Although PST Nm was able to add PSA to NCAM, most of its product was attached to other cell surface proteins. Nevertheless, the PST Nm-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 PST Nm 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 PST Nm-based approach may provide a valuable alternative to PST gene therapy. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc.
Keywords: controlled study; unclassified drug; drug activity; nonhuman; animal cell; mouse; animals; mice; animal tissue; gene expression; embryonic stem cell; animal experiment; brain cortex; in vivo study; neurons; in vitro study; bacterial proteins; cell culture; brain; spinal cord; polysialic acid; rat; sialyltransferase; sialyltransferases; cell types; cell membranes; cell migration; fibroblast; in-vivo; rats; schwann cell; cerebral cortex; in-vitro; cell adhesion; tissue; corpus striatum; sialic acids; chickens; rats, inbred f344; cho cell; cho cells; cricetinae; cricetulus; neural cell adhesion molecules; spinal cords; chicken; purification; neurite outgrowth; adult brain; nerve fiber growth; cellular process; neisseria meningitidis; cell engineering; striatum; polysialylation; cell surface proteins; chinese hamster ovary cells; donor substrates; extracellular environments; mouse embryonic stem cells; rat schwann cell; stx genes; polysialyltransferase; adult animal; enzyme engineering; metabolic engineering
Journal Title: Journal of Biological Chemistry
Volume: 287
Issue: 39
ISSN: 0021-9258
Publisher: American Society for Biochemistry and Molecular Biology  
Date Published: 2012-09-21
Start Page: 32770
End Page: 32779
Language: English
DOI: 10.1074/jbc.M112.377614
PROVIDER: scopus
PMCID: PMC3463313
PUBMED: 22851175
DOI/URL:

http://www.scopus.com/inward/record.url?eid=2-s2.0-84866534476&partnerID=40&md5=2ad2b1dd9a8e5c9c66cec2efcb077ec0
Notes: --- - "Export Date: 2 November 2012" - "CODEN: JBCHA" - "Source: Scopus"
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MSK Authors
  1. Urs S Rutishauser
    70 Rutishauser
  2. Abderrahman El Maarouf
    22 El Maarouf
  3. Damali Moyo-Lee Yaw
    2 Moyo-Lee Yaw
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