Biochemical characterization of a polysialyltransferase from Mannheimia haemolytica A2 and comparison to other bacterial polysialyltransferases

Download
  1. Get@NRC: Biochemical characterization of a polysialyltransferase from Mannheimia haemolytica A2 and comparison to other bacterial polysialyltransferases (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1371/journal.pone.0069888
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titlePLoS ONE
ISSN1932-6203
Volume8
Issue7
Article numbere69888
Subjectbacterial enzyme; glycan; glycoprotein; hybrid protein; maltose binding protein; polysialic acid; polysialyltransferase; recombinant enzyme; unclassified drug; article; binding site; carbohydrate analysis; cell migration; cell surface; comparative study; DNA sequence; enzyme activity; enzyme analysis; enzyme kinetics; enzyme stability; enzyme synthesis; Escherichia coli; Mannheimia haemolytica; Neisseria meningitidis; neuroprogenitor cell; nonhuman; nucleotide sequence; pH; protein expression; protein localization; protein modification; solubility; stem cell; suicide substrate; thermostability; wound healing
AbstractPolysialic acids are bioactive carbohydrates found in eukaryotes and some bacterial pathogens. The bacterial polysialyltransferases (PSTs), which catalyze the synthesis of polysialic acid capsules, have previously been identified in select strains of Escherichia coli and Neisseria meningitidis and are classified in the Carbohydrate-Active enZYmes Database as glycosyltransferase family GT-38. In this study using DNA sequence analysis and functional characterization we have identified a novel polysialyltransferase from the bovine/ovine pathogen Mannheimia haemolytica A2 (PSTMh). The enzyme was expressed in recombinant form as a soluble maltose-binding-protein fusion in parallel with the related PSTs from E. coli K1 and N. meningitidis group B in order to perform a side-by-side comparison. Biochemical properties including solubility, acceptor preference, reaction pH optima, thermostability, kinetics, and product chain length for the enzymes were compared using a synthetic fluorescent acceptor molecule. PSTMh exhibited biochemical properties that make it an attractive candidate for chemi-enzymatic synthesis applications of polysialic acid. The activity of PSTMh was examined on a model glycoprotein and the surface of a neuroprogenitor cell line where the results supported its development for use in applications to therapeutic protein modification and cell surface glycan remodelling to enable cell migration at implantation sites to promote wound healing. The three PSTs examined here demonstrated different properties that would each be useful to therapeutic applications. © 2013 Lindhout et al.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada (NRC-CNRC); Human Health Therapeutics (HHT-TSH)
Peer reviewedYes
NPARC number21269716
Export citationExport as RIS
Report a correctionReport a correction
Record identifier8e519624-1cfd-4675-bfa0-d3f56edf563d
Record created2013-12-13
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)