Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins.
The aim of this study was to test whether galectin-3 is present in human corneal epithelium and whether lipopolysaccharide (LPS) purified from Pseudomonas aeruginosa ATCC 19660 binds to this animal lectin and/or to another human corneal epithelial protein(s) (HCEP) and to confirm which component of LPS (inner or outer core or lipid A) is important in bacterial binding by using the eye in organ culture. LPS isolated and purified from P. aeruginosa ATCC 19660 and a commercial LPS (serotype 10) differed in polyacrylamide gel analysis but bound similarly to blotted HCEP. Binding was determined to be a receptor-ligand type of interaction by the solid-phase assay, because it was both specific and saturable. Several LPS binding proteins in HCEP were identified by an overlay method. Western blotting with antibody against galectin-3 revealed the presence of this protein in both freshly isolated and cultured transformed human corneal epithelium. Binding inhibition assays showed that antibody specific for the outer core region of LPS and an anti-galectin antibody significantly inhibited bacterial binding in vitro. These data provide further evidence that LPS is an important adhesin of P. aeruginosa, that it binds to protein receptor molecules in HCEP, that one of the LPS binding proteins is galectin-3, and that the outer core portion of the molecule appears to be critical for LPS binding to the eye.
Gupta
S K
SK
Department of Anatomy/Cell Biology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.
Masinick
S
S
Garrett
M
M
Hazlett
L D
LD
D000943:Antigens, Differentiation / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D002478:Cells, Cultured; D003315:Cornea / Q000737:chemistry* / Q000276:immunology; D004848:Epithelium / Q000737:chemistry / Q000276:immunology; D005136:Eye Proteins / Q000276:immunology / Q000378:metabolism*; D037502:Galectin 3; D006801:Humans; D008070:Lipopolysaccharides / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000276:immunology / Q000378:metabolism*
D000943:Antigens, Differentiation; D005136:Eye Proteins; D037502:Galectin 3; D008070:Lipopolysaccharides