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A Novel secreted endoglycosidase from Enterococcus faecalis with activity on human immunoglobulin G and ribonuclease B

Collin, Mattias LU and Fischetti, Vincent A. (2004) In Journal of Biological Chemistry 279(21). p.22558-22570
Abstract

The human pathogen Enterococcus faecalis can degrade the N-linked glycans of human RNase B to acquire nutrients, but no gene or protein has been associated with this activity. We identified an 88-kDa secreted protein, endoglycosidase (Endo) E, which is most likely responsible for this activity. EndoE, encoded by ndoE, consists of an α-domain with a family 18 glycosyl hydrolase motif and a β-domain similar to family 20 glycosyl hydrolases. Phylogenetic analysis of EndoE indicates that the α-domain is related to human chitobiases, and the β-domain is related to bacterial and human hexosaminidases. Recombinant expression of full-length EndoE or EndoEα, site-directed mutagenesis of the catalytic residues, mass spectroscopy, and homology... (More)

The human pathogen Enterococcus faecalis can degrade the N-linked glycans of human RNase B to acquire nutrients, but no gene or protein has been associated with this activity. We identified an 88-kDa secreted protein, endoglycosidase (Endo) E, which is most likely responsible for this activity. EndoE, encoded by ndoE, consists of an α-domain with a family 18 glycosyl hydrolase motif and a β-domain similar to family 20 glycosyl hydrolases. Phylogenetic analysis of EndoE indicates that the α-domain is related to human chitobiases, and the β-domain is related to bacterial and human hexosaminidases. Recombinant expression of full-length EndoE or EndoEα, site-directed mutagenesis of the catalytic residues, mass spectroscopy, and homology modeling shows that EndoEα hydrolyzes the glycan on human RNase B, whereas EndoEβ hydrolyzes the conserved glycan on IgG. Denaturation experiments indicate that the chitinase activity on RNase B is not dependent on the tertiary structure, although it is on IgG. The ndoE gene and secreted EndoE are present in most E. faecalis but not in Enterococcus faecium isolates. Correspondingly, E. faecalis, but not E. faecium, degrades the glycan on RNase B during growth. Thus, we have identified a secreted enzyme from E. faecalis, EndoE, which by two distinct activities hydrolyzes the glycans on RNase B and IgG. Both activities could be important for the molecular pathogenesis and persistence of E. faecalis during human infections.

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author
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publication status
published
subject
in
Journal of Biological Chemistry
volume
279
issue
21
pages
13 pages
publisher
ASBMB
external identifiers
  • scopus:2542467597
ISSN
0021-9258
DOI
10.1074/jbc.M402156200
language
English
LU publication?
no
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ff924790-13d7-46a0-b93c-6899e0d1dc9f
date added to LUP
2018-10-30 19:48:29
date last changed
2018-11-21 21:42:53
@article{ff924790-13d7-46a0-b93c-6899e0d1dc9f,
  abstract     = {<p>The human pathogen Enterococcus faecalis can degrade the N-linked glycans of human RNase B to acquire nutrients, but no gene or protein has been associated with this activity. We identified an 88-kDa secreted protein, endoglycosidase (Endo) E, which is most likely responsible for this activity. EndoE, encoded by ndoE, consists of an α-domain with a family 18 glycosyl hydrolase motif and a β-domain similar to family 20 glycosyl hydrolases. Phylogenetic analysis of EndoE indicates that the α-domain is related to human chitobiases, and the β-domain is related to bacterial and human hexosaminidases. Recombinant expression of full-length EndoE or EndoEα, site-directed mutagenesis of the catalytic residues, mass spectroscopy, and homology modeling shows that EndoEα hydrolyzes the glycan on human RNase B, whereas EndoEβ hydrolyzes the conserved glycan on IgG. Denaturation experiments indicate that the chitinase activity on RNase B is not dependent on the tertiary structure, although it is on IgG. The ndoE gene and secreted EndoE are present in most E. faecalis but not in Enterococcus faecium isolates. Correspondingly, E. faecalis, but not E. faecium, degrades the glycan on RNase B during growth. Thus, we have identified a secreted enzyme from E. faecalis, EndoE, which by two distinct activities hydrolyzes the glycans on RNase B and IgG. Both activities could be important for the molecular pathogenesis and persistence of E. faecalis during human infections.</p>},
  author       = {Collin, Mattias and Fischetti, Vincent A.},
  issn         = {0021-9258},
  language     = {eng},
  month        = {05},
  number       = {21},
  pages        = {22558--22570},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {A Novel secreted endoglycosidase from Enterococcus faecalis with activity on human immunoglobulin G and ribonuclease B},
  url          = {http://dx.doi.org/10.1074/jbc.M402156200},
  volume       = {279},
  year         = {2004},
}