Isolation and characterisation of putative adhesins from Helicobacter pylori with affinity for heparan sulphate proteoglycan.
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-PAGE 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.
Ruiz-Bustos
E
E
Ochoa
J L
JL
Wadström
T
T
Department of Marine Pathology, Center for Biological Research, La Paz, Baja California Sur, 23000 México and *University of Lund, Institute of Medical Microbiology, Sölvegatan 23, S-223 62 Lund, Sweden.
Ascencio
F
F
D018829:Adhesins, Bacterial / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D002352:Carrier Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D016480:Helicobacter pylori / Q000502:physiology*; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D007525:Isoelectric Focusing; D008970:Molecular Weight
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D002352:Carrier Proteins; D019812:Heparan Sulfate Proteoglycans; C073899:adhesin, helicobacter