Journal article
Specificity of the high-mannose recognition site between Enterobacter cloacae pili adhesin and HT-29 cell membranes
Infection and immunity, Vol.65(10), pp.4199-4206
10/1997
DOI: 10.1128/iai.65.10.4199-4206.1997
PMCID: PMC175603
PMID: 9317027
Abstract
Enterobacter cloacae has been implicated as one of the causative agents in neonatal infection and causes a septicemia thought to be initiated via the gastrointestinal tract. The adhesion of radiolabeled E. cloacae to HT-29 cells was concentration and temperature dependent and was effectively blocked by unlabeled bacteria or by millimolar concentrations of alpha-mannosides and micromolar concentrations of high-mannose oligosaccharides. A variety of well-characterized mannose oligosaccharides were tested as inhibitors of adhesion. The best inhibitor was the Man9(GlcNAc)2-tyrosinamide, which was considerably better than other tyrosinamide-linked oligosaccharides such as Man7(GlcNAc)2, Man6(GlcNAc)2 or Man5(GlcNAc)2. Further evidence that the bacteria preferred Man9(GlcNAc)2 structures was obtained by growing HT-29 cells in the presence of glycoprotein processing inhibitors that block mannosidase I and increase the amount of protein-bound Man9(GlcNAc)2 at the cell surface. Such cells bound 1.5- to 2-fold more bacteria than did control cells. The adhesin involved in binding to high-mannose structures was purified from isolated pili. On sodium dodecyl sulfate-gels, a 35-kDa protein was identified by its specific binding to a mannose-containing biotinylated albumin. The amino acid sequences of several peptides from the 35-kDa subunit showed over 85% identity to FimH, the mannose-specific adhesin of Salmonella typhimurium. Pili were labeled with 125I and examined for the ability to bind to HT-29 cells. Binding showed saturation kinetics and was inhibited by the addition of Man9(GlcNAc)2-tyrosinamide but not by oligosaccharides with fewer mannose residues. Polyclonal antibody against this 35-kDa protein also effectively blocked adhesion of pili or E. cloacae, but no effect was observed with nonspecific antibody. These studies demonstrate that the 35-kDa pilus subunit is a lectin whose specificity is directed toward Man, (GlcNAc)2 oligosaccharides.
Details
- Title: Subtitle
- Specificity of the high-mannose recognition site between Enterobacter cloacae pili adhesin and HT-29 cell membranes
- Creators
- Y T Pan - University of Arkansas for Medical SciencesBin Xu - University of Arkansas for Medical SciencesKevin Rice - University of Arkansas for Medical SciencesSam Smith - University of Arkansas for Medical SciencesRichard Jackson - University of Arkansas for Medical SciencesAlan D Elbein - University of Arkansas for Medical Sciences
- Resource Type
- Journal article
- Publication Details
- Infection and immunity, Vol.65(10), pp.4199-4206
- DOI
- 10.1128/iai.65.10.4199-4206.1997
- PMID
- 9317027
- PMCID
- PMC175603
- NLM abbreviation
- Infect Immun
- ISSN
- 0019-9567
- eISSN
- 1098-5522
- Language
- English
- Date published
- 10/1997
- Academic Unit
- Pharmaceutical Sciences and Experimental Therapeutics; Craniofacial Anomalies Research Center; Medicinal and Natural Products Chemistry
- Record Identifier
- 9984365903102771
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