Proteinase K decreases Pseudomonas aeruginosa adhesion to wounded cornea.
The present study was designed to obtain further information regarding the molecular nature of the corneal receptor(s) facilitating Pseudomonas aeruginosa adhesion to cornea. Scarified adult mouse corneas in organ culture were treated for 10 or 60 min with a panel of lipase-free proteases [each at 20 micrograms ml-1 or 0.22 Units (U) ml-1, activity] including trypsin, chymotrypsin, V8 protease, elastase, subtilisin A, pronase protease and proteinase K. All of these, except proteinase K treatment (20 micrograms ml-1 for 60 min), either significantly elevated or had no effect (proteinase K 20 micrograms ml-1 for 10 min) on subsequent bacterial adhesion at 60 min following topical application of the inoculum to the scarified corneal surface. Enzyme treatment times of 10, 30 or 60 min at a higher concentration (50 micrograms ml-1) of proteinase K, significantly decreased binding at 60 min after bacterial application for each enzyme treatment time. The combined effects of proteases and lipase on bacterial binding also was examined. Eyes treated with proteinase K (20 micrograms ml-1 for 1 hr) or protease-free lipase (50,000 U ml-1 for 1 hr) alone or in combination, all reduced bacterial binding, but the effect was not additive. Trypsin or lipase alone significantly enhanced or decreased binding, respectively. In contrast, trypsin (20 micrograms ml-1 for 1 hr) followed by lipase treatment (50,000 U ml-1 for 1 hr) resulted in binding which was not significantly different than phosphate-buffered saline (PBS) control binding, indicating that the trypsin exposed receptor was lipase sensitive.(ABSTRACT TRUNCATED AT 250 WORDS)
Hazlett
L D
LD
Department of Anatomy/Cell Biology, Wayne State University, Detroit, MI 48201.
Masinick
S
S
Berk
R S
RS
Zheng
Z
Z
D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects*; D003315:Cornea / Q000382:microbiology / Q000648:ultrastructure; D065306:Corneal Injuries; D019286:Endopeptidase K; D010450:Endopeptidases / Q000494:pharmacology; D005260:Female; D008049:Lipase / Q000494:pharmacology; D051379:Mice; D008813:Mice, Inbred ICR; D008855:Microscopy, Electron, Scanning; D009924:Organ Culture Techniques; D011550:Pseudomonas aeruginosa; D012697:Serine Endopeptidases / Q000494:pharmacology*
D008049:Lipase; D010450:Endopeptidases; D012697:Serine Endopeptidases; D019286:Endopeptidase K