title: Corneal surface glycosylation is modulated by IL-1R andPseudomonas aeruginosachallenge but is insufficient for inhibiting bacterial binding.
abstract: Cell surface glycosylation is thought to be involved in barrier function against microbes at mucosal surfaces. Previously we showed that the epithelium of healthy mouse corneas becomes vulnerable toPseudomonas aeruginosaadhesion if it lacks the innate defense protein MyD88 (myeloid differentiation primary response gene 88), or after superficial injury by blotting with tissue paper. Here we explored their effect on corneal surface glycosylation using a metabolic label, tetra-acetylatedN-azidoacetylgalactosamine (Ac4GalNAz). Ac4GalNAz treatment labeled the surface of healthy mouse corneas, leaving most cells viable, and bacteria preferentially associated with GalNAz-labeled regions. Surprisingly, corneas from MyD88-/-mice displayed similar GalNAz labeling to wild-type corneas, but labeling was reduced and patchy on IL-1 receptor (IL-1R)-knockout mouse corneas (P< 0.05, ANOVA). Tissue paper blotting removed GalNAz-labeled surface cells, causing DAPI labeling (permeabilization) of underlying cells. MS of material collected on the tissue paper blots revealed 67 GalNAz-labeled proteins, including intracellular proteins. These data show that the normal distribution of surface glycosylation requires IL-1R, but not MyD88, and is not sufficient to prevent bacterial binding. They also suggest increasedP. aeruginosaadhesion to MyD88-/-and blotted corneas is not due to reduction in total surface glycosylation, and for tissue paper blotting is likely due to cell permeabilization.-Jolly, A. L., Agarwal, P., Metruccio, M. M. E., Spiciarich, D. R., Evans, D. J., Bertozzi, C. R., Fleiszig, S. M. J. Corneal surface glycosylation is modulated by IL-1R andPseudomonas aeruginosachallenge but is insufficient for inhibiting bacterial binding.
authors.affiliation: School of Optometry, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Amber L
authors.initials: AL
authors.lastname: Jolly
authors.affiliation: College of Chemistry, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Paresh
authors.initials: P
authors.lastname: Agarwal
authors.affiliation: School of Optometry, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Matteo M E
authors.initials: MME
authors.lastname: Metruccio
authors.affiliation: College of Chemistry, University of California, Berkeley, Berkeley, California, USA.
authors.forename: David R
authors.initials: DR
authors.lastname: Spiciarich
authors.affiliation: School of Optometry, University of California, Berkeley, Berkeley, California, USA.|College of Pharmacy, Touro University California, Vallejo, California, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Evans
authors.affiliation: College of Chemistry, University of California, Berkeley, Berkeley, California, USA.|Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.|Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Carolyn R
authors.initials: CR
authors.lastname: Bertozzi
authors.affiliation: School of Optometry, University of California, Berkeley, Berkeley, California, USA; fleiszig@berkeley.edu.|Graduate Division of Vision Sciences, University of California, Berkeley, Berkeley, California, USA.|Graduate Division of Microbiology, University of California, Berkeley, Berkeley, California, USA.|Graduate Division of Infectious Diseases and Immunity, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Suzanne M J
authors.initials: SMJ
authors.lastname: Fleiszig
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D003315:Cornea / Q000382:microbiology* / Q000502:physiology*; D005260:Female; D015718:Fibrin Tissue Adhesive; D005786:Gene Expression Regulation / Q000502:physiology; D006023:Glycoproteins; D006031:Glycosylation; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000378:metabolism*; D011550:Pseudomonas aeruginosa; D017472:Receptors, Interleukin-1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D015718:Fibrin Tissue Adhesive; D006023:Glycoproteins; C507780:Myd88 protein, mouse; D053594:Myeloid Differentiation Factor 88; D017472:Receptors, Interleukin-1
keywords: GalNAz metabolic label; Muc4; MyD88; imaging; mass spectrometry