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Characterization and Acceptor Preference of a Soluble Meningococcal Group C Polysialyltransferase

Peterson, Dwight C and Arakere, Gayathri and Vionnet, Justine and McCarthy, Pumtiwitt C and Vann, Willie F (2011) Characterization and Acceptor Preference of a Soluble Meningococcal Group C Polysialyltransferase. In: Journal of Bacteriology, 193 (7). pp. 1576-1582.

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Official URL: http://jb.asm.org/cgi/content/abstract/193/7/1576

Abstract

Vaccines against Neisseria meningitidis group C are based on its alpha-2,9-linked polysialic acid capsular polysaccharide. This polysialic acid expressed on the surface of N. meningitidis and in the absence of specific antibody serves to evade host defense mechanisms. The polysialyltransferase (PST) that forms the group C polysialic acid (NmC PST) is located in the cytoplasmic membrane. Until recently, detailed characterization of bacterial polysialyltransferases has been hampered by a lack of availability of soluble enzyme preparations. We have constructed chimeras of the group C polysialyltransferase that catalyzes the formation alpha-2,9-polysialic acid as a soluble enzyme. We used site-directed mutagenesis to determine the region of the enzyme necessary for synthesis of the alpha-2,9 linkage. A chimera of NmB and NmC PSTs containing only amino acids 1 to 107 of the NmB polysialyltransferase catalyzed the synthesis of alpha-2,8-polysialic acid. The NmC polysialyltransferase requires an exogenous acceptor for catalytic activity. While it requires a minimum of a disialylated oligosaccharide to catalyze transfer, it can form high-molecular-weight alpha-2,9-polysialic acid in a nonprocessive fashion when initiated with an alpha-2,8-polysialic acid acceptor. De novo synthesis in vivo requires an endogenous acceptor. We attempted to reconstitute de novo activity of the soluble group C polysialyltransferase with membrane components. We found that an acapsular mutant with a defect in the polysialyltransferase produces outer membrane vesicles containing an acceptor for the alpha-2,9-polysialyltransferase. This acceptor is an amphipathic molecule and can be elongated to produce polysialic acid that is reactive with group C-specific antibody.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to American Society for Microbiology.
Department/Centre: Others
Date Deposited: 08 Apr 2011 07:24
Last Modified: 08 Apr 2011 07:24
URI: http://eprints.iisc.ernet.in/id/eprint/36573

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