Identification of outer membrane Porin D as a vitronectin-binding factor in cystic fibrosis clinical isolates of Pseudomonas aeruginosa

J Cyst Fibros. 2015 Sep;14(5):600-7. doi: 10.1016/j.jcf.2015.05.005. Epub 2015 Jun 3.

Abstract

Background: Pseudomonas aeruginosa is a pathogen that frequently colonizes patients with cystic fibrosis (CF) or chronic obstructive pulmonary disease (COPD). Several pathogens are known to bind vitronectin to increase their virulence. Vitronectin has been shown to enhance P. aeruginosa adhesion to host epithelial cells.

Methods: We screened clinical isolates from the airways of CF patients and from the bloodstream of patients with bacteremia for binding of vitronectin. Two-dimensional SDS-PAGE and a proteomic approach were used to identify vitronectin-receptors in P. aeruginosa.

Results: P. aeruginosa from the airways of CF patients (n=27) bound more vitronectin than bacteremic isolates (n=15, p=0.025). Porin D (OprD) was identified as a vitronectin-binding protein. A P. aeruginosa oprD transposon insertion mutant had a decreased binding to soluble and immobilized vitronectin (p≤0.001).

Conclusions: P. aeruginosa isolates obtained from CF patients significantly bound vitronectin. Porin D was defined as a novel P. aeruginosa vitronectin-receptor, and we postulate that the Porin D-dependent interaction with vitronectin may be important for colonization.

Keywords: Adhesion molecules; Extracellular matrix; Porin D; Pseudomonas aeruginosa; Vitronectin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Outer Membrane Proteins / biosynthesis
  • Bacterial Outer Membrane Proteins / genetics
  • Blotting, Western
  • Cystic Fibrosis / genetics
  • Cystic Fibrosis / metabolism
  • Cystic Fibrosis / microbiology*
  • DNA, Bacterial / genetics*
  • Disease Models, Animal
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Expression Regulation, Bacterial*
  • Humans
  • Porins / biosynthesis
  • Porins / genetics*
  • Pseudomonas Infections / genetics
  • Pseudomonas Infections / metabolism
  • Pseudomonas Infections / microbiology*
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / isolation & purification*
  • Rabbits
  • Respiratory System / microbiology
  • Vitronectin / biosynthesis
  • Vitronectin / genetics*

Substances

  • Bacterial Outer Membrane Proteins
  • DNA, Bacterial
  • Porins
  • Vitronectin
  • OprD protein, Pseudomonas aeruginosa