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Isolation and characterisation of putative adhesins from Helicobacter pylori with affinity for heparan sulphate proteoglycan

Ruiz-Bustos, E.; Ochoa, J.L.; Wadström, Torkel LU and Ascencio, F. (2001) In Journal of Medical Microbiology 50(3). p.215-222
Abstract
A pool of heparan sulphate-binding proteins (HSBPs) from Helicobacter pylori culture supernates was obtained by sequential ammonium sulphate precipitation and affinity chromatography on heparin-Sepharose, The chromatographic procedure yielded one major fraction that contained proteins with heparan sulphate affinity as revealed by inhibition studies of heparan sulphate binding to H. pylori cells. Preparative iso-electric focusing, SDS-PACE and blotting experiments, with peroxidase(POD)-labelled heparan sulphate as a probe, indicated the presence of two major extracellular proteins with POD-heparan sulphate affinity. One protein had a molecular mass of 66.2 kDa and a pI of 5.4, whilst the second protein had a molecular mass of 71.5 kDa and a... (More)
A pool of heparan sulphate-binding proteins (HSBPs) from Helicobacter pylori culture supernates was obtained by sequential ammonium sulphate precipitation and affinity chromatography on heparin-Sepharose, The chromatographic procedure yielded one major fraction that contained proteins with heparan sulphate affinity as revealed by inhibition studies of heparan sulphate binding to H. pylori cells. Preparative iso-electric focusing, SDS-PACE and blotting experiments, with peroxidase(POD)-labelled heparan sulphate as a probe, indicated the presence of two major extracellular proteins with POD-heparan sulphate affinity. One protein had a molecular mass of 66.2 kDa and a pI of 5.4, whilst the second protein had a molecular mass of 71.5 kDa and a pI of 5.0. The N-terminal amino acid sequence of the 71.5-kDa HSBP did not show homology to any other heparin-binding protein, nor to known proteins of H, pylori, whereas the 66.2-kDa HSBP showed a high homology to an Escherichia coli chaperon protein and equine haemoglobin. A third HSBP was isolated from an outer-membrane protein (OMP) fraction of H. pylori cells with a molecular mass of 47.2 kDa, The amino acid sequence of an internal peptide of the OMP-HSBP did not show homology to the extracellular HSBP of H, pylori, or to another microbial HSBP. (Less)
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Contribution to journal
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published
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in
Journal of Medical Microbiology
volume
50
issue
3
pages
215 - 222
publisher
Lippincott Williams & Wilkins
external identifiers
  • wos:000167047100003
  • scopus:0035113648
ISSN
0022-2615
language
English
LU publication?
yes
id
99a243fd-cc1e-477b-9666-f44509b99c62 (old id 1119565)
alternative location
http://jmm.sgmjournals.org/cgi/content/abstract/50/3/215
date added to LUP
2008-07-14 08:52:55
date last changed
2018-01-07 09:51:44
@article{99a243fd-cc1e-477b-9666-f44509b99c62,
  abstract     = {A pool of heparan sulphate-binding proteins (HSBPs) from Helicobacter pylori culture supernates was obtained by sequential ammonium sulphate precipitation and affinity chromatography on heparin-Sepharose, The chromatographic procedure yielded one major fraction that contained proteins with heparan sulphate affinity as revealed by inhibition studies of heparan sulphate binding to H. pylori cells. Preparative iso-electric focusing, SDS-PACE and blotting experiments, with peroxidase(POD)-labelled heparan sulphate as a probe, indicated the presence of two major extracellular proteins with POD-heparan sulphate affinity. One protein had a molecular mass of 66.2 kDa and a pI of 5.4, whilst the second protein had a molecular mass of 71.5 kDa and a pI of 5.0. The N-terminal amino acid sequence of the 71.5-kDa HSBP did not show homology to any other heparin-binding protein, nor to known proteins of H, pylori, whereas the 66.2-kDa HSBP showed a high homology to an Escherichia coli chaperon protein and equine haemoglobin. A third HSBP was isolated from an outer-membrane protein (OMP) fraction of H. pylori cells with a molecular mass of 47.2 kDa, The amino acid sequence of an internal peptide of the OMP-HSBP did not show homology to the extracellular HSBP of H, pylori, or to another microbial HSBP.},
  author       = {Ruiz-Bustos, E. and Ochoa, J.L. and Wadström, Torkel and Ascencio, F.},
  issn         = {0022-2615},
  language     = {eng},
  number       = {3},
  pages        = {215--222},
  publisher    = {Lippincott Williams & Wilkins},
  series       = {Journal of Medical Microbiology},
  title        = {Isolation and characterisation of putative adhesins from Helicobacter pylori with affinity for heparan sulphate proteoglycan},
  volume       = {50},
  year         = {2001},
}