identification of a human lactoferrin binding protein in gardnerella vaginalis by jacinta gaitor l.
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Identification of a Human Lactoferrin-Binding Protein in Gardnerella vaginalis. By Jacinta Gaitor. Jarosik, G et al. 2000. Identification of a Human Lactoferrin-Binding protein in Gardnerella vaginalis. Infection and immunity. p. 3443-3447. Introduction.

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identification of a human lactoferrin binding protein in gardnerella vaginalis by jacinta gaitor

Identification of a Human Lactoferrin-Binding Protein in Gardnerella vaginalis.By Jacinta Gaitor

Jarosik, G et al. 2000. Identification of a Human Lactoferrin-Binding protein in Gardnerella vaginalis. Infection and immunity. p. 3443-3447

introduction
Introduction
  • Gardnerella vaginalis is associated with Bacterial vaginosis
  • Change in vaginal microflora
  • Lactobacillus vs anaerobes and Gardnerella Vaginalis
  • Upper urinary tract infections
  • Preterm delivery
  • PROM
  • Increased risk of HIV virus and transmission
gardnerella vaginalis
Gardnerella vaginalis
  • Non-motile
  • Beta hemolytic
  • Unencapsulated
  • Rod shaped
  • Gram positive cell wall
  • Pilli
  • Exopolysaccharide coat
iron and its relation to microrganisms
Iron and its relation to microrganisms
  • Necessity
  • Heme, ferritin, hemoglobin
  • Transferrin, lactoferrin
lactoferrin
Lactoferrin
  • Iron binding protein
  • Kd 10^-19
  • pH 6.4
  • 26-folds greater than transferrin
  • Found in milk, neutrophils leukocytes
mechanism of obtaining iron
Mechanism of Obtaining Iron
  • Siderophores
  • Direct binding of iron containing compounds by cell receptors
  • Hemolysins and cytolysins
gardnerella and iron
Gardnerella and Iron
  • Iron salts, heme, hemoglobin, Lactoferrin
  • Human Lactoferrin (hLf)
the binding of g vaginalis and hlf
The Binding of G. vaginalis and hLf
  • G. vaginalis 594 and 317 and E. coli
  • DIG-hLf probe
  • Chemiluminescence detection
protein involved in the binding of dig hlf
Protein Involved in the Binding of DIG - hLf
  • Western block analysis
  • Proteins from G. vaginalis lysates seperated by SDS – PAGE
  • DIG –hLf probe
  • ~120kDA Lbp
detection of 120kda lbp under different iron circumstances
Detection of 120kDA Lbp Under Different Iron Circumstances
  • G.vaginalis 317
  • Iron replete conditions
  • Iron supplemented conditions
  • Iron deficient conditions
  • Iron activity can be regulated
specificity of the 120kda lbp binding acivity
Specificity of the 120kDaLbp binding acivity
  • Unlabelled preincubation iron compounds
  • Unlabelled hLf inhibited binding of DIG-hLf by 120kDa protein
  • Binding acivity for 120kDA Lbp specific for hLf
proteolytic treatment of g vaginalis cells
Proteolytic treatment of G. vaginalis cells
  • G. vaginalis 317 cells
  • Proteinase K or Trypsin
  • Binding of DIG-hLf found among cells not treated with protease.
  • 120kDa protein may be surface exposed
conclusion
Conclusion
  • hLf – extracellular iron-binding glycoprotein
  • G.vaginalis can utilize hLf as a sole source of iron
  • G. vagionalis Lbp specific for hLf
  • Surface exposed
references
References
  • The Dictionary of Cell and Molecular Biology.http://www.mblab.gla.ac.uk/~julian/Dict.html
  • Jarosik, G et al. 2000. Identification of a Human Lactoferrin-Binding protein in Gardnerella vaginalis. Infection and immunity. p. 3443-3447