PMID- 23922842
OWN - NLM
STAT- MEDLINE
DCOM- 20140429
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 7
DP  - 2013
TI  - Biochemical characterization of a polysialyltransferase from Mannheimia 
      haemolytica A2 and comparison to other bacterial polysialyltransferases.
PG  - e69888
LID - 10.1371/journal.pone.0069888 [doi]
LID - e69888
AB  - Polysialic 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.
FAU - Lindhout, Theresa
AU  - Lindhout T
AD  - Department of Molecular and Cellular Biology, University of Guelph, Guelph, 
      Ontario, Canada.
FAU - Bainbridge, Cynthia R
AU  - Bainbridge CR
FAU - Costain, Will J
AU  - Costain WJ
FAU - Gilbert, Michel
AU  - Gilbert M
FAU - Wakarchuk, Warren W
AU  - Wakarchuk WW
LA  - eng
GR  - MOP-84272/Canadian Institutes of Health Research/Canada
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130726
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Bacterial Proteins)
RN  - 0 (Fetuins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Sialic Acids)
RN  - 0 (cytidine-5'-diphosphosialic acid)
RN  - 0 (polysialic acid)
RN  - 63-38-7 (Cytidine Diphosphate)
RN  - EC 2.4.99.- (Sialyltransferases)
SB  - IM
MH  - Animals
MH  - Bacterial Proteins/metabolism
MH  - Cell Membrane/metabolism
MH  - Cytidine Diphosphate/analogs & derivatives/metabolism
MH  - Electrophoresis, Capillary
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Enzyme Stability
MH  - Escherichia coli/*enzymology
MH  - Fetuins/metabolism
MH  - Genome, Bacterial/genetics
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Mannheimia haemolytica/*enzymology/genetics
MH  - Neisseria meningitidis, Serogroup B/*enzymology
MH  - PC12 Cells
MH  - Rats
MH  - Recombinant Proteins/metabolism
MH  - Sialic Acids/metabolism
MH  - Sialyltransferases/*metabolism
MH  - Solubility
MH  - Temperature
MH  - Time Factors
PMC - PMC3724679
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/08/08 06:00
MHDA- 2014/04/30 06:00
PMCR- 2013/07/26
CRDT- 2013/08/08 06:00
PHST- 2013/03/27 00:00 [received]
PHST- 2013/06/13 00:00 [accepted]
PHST- 2013/08/08 06:00 [entrez]
PHST- 2013/08/08 06:00 [pubmed]
PHST- 2014/04/30 06:00 [medline]
PHST- 2013/07/26 00:00 [pmc-release]
AID - PONE-D-13-12726 [pii]
AID - 10.1371/journal.pone.0069888 [doi]
PST - epublish
SO  - PLoS One. 2013 Jul 26;8(7):e69888. doi: 10.1371/journal.pone.0069888. Print 2013.