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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.

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

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  1. 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

  2. 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

  3. Gardnerella vaginalis • Non-motile • Beta hemolytic • Unencapsulated • Rod shaped • Gram positive cell wall • Pilli • Exopolysaccharide coat

  4. Iron and its relation to microrganisms • Necessity • Heme, ferritin, hemoglobin • Transferrin, lactoferrin

  5. Lactoferrin • Iron binding protein • Kd 10^-19 • pH 6.4 • 26-folds greater than transferrin • Found in milk, neutrophils leukocytes

  6. Mechanism of Obtaining Iron • Siderophores • Direct binding of iron containing compounds by cell receptors • Hemolysins and cytolysins

  7. Gardnerella and Iron • Iron salts, heme, hemoglobin, Lactoferrin • Human Lactoferrin (hLf)

  8. The Binding of G. vaginalis and hLf • G. vaginalis 594 and 317 and E. coli • DIG-hLf probe • Chemiluminescence detection

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. 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

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