title: Identification of a high-mannose ICAM-1 glycoform: effects of ICAM-1 hypoglycosylation on monocyte adhesion and outside in signaling.
abstract: Endothelial adhesion molecules are critical effectors of inflammation ensuring coordinated interactions that allow leukocytes to home to sites of injury. These adhesion molecules are often extensively modified posttranslationaly by the addition of N-glycans, but if, or how, these modifications contribute to the protein function remains poorly understood. Herein we show that activated endothelial cells express two distinct N-glycoforms of intercellular adhesion molecule 1 (ICAM-1) that comprise a complex N-glycoform with α-2,6 sialic acid present at relatively high levels and a second, less abundant and previously undescribed high-mannose glycoform (HM-ICAM-1). This novel HM-ICAM-1 glycoform was also detected in human coronary artery specimens and moreover appeared to be the dominant glycoform in vivo. Production of exclusively HM-ICAM-1 in cells by α-mannosidase inhibition increased monocyte rolling and adhesion compared with mature ICAM-1 consistent with high-mannose epitopes providing leukocyte ligands. Cross-linking of ICAM-1 transmits outside-in signals that affect endothelial permeability and survival. Interestingly, cell signaling (assessed using ERK, VE-cadherin, and Akt phosphorylation) was maintained after cross-linking of HM-ICAM-1 compared with mature ICAM-1; however, interactions with the actin cytoskeleton were lost with HM-ICAM-1. These findings suggest that specific ICAM-1 N-glycoforms modulate distinct aspects of the inflammatory response and identify HM-ICAM-1 as a new therapeutic target for controlling leukocyte trafficking and endothelial inflammation.
authors.affiliation: Department of Pathology, University of Alabama, Birmingham, Alabama 35294, USA.
authors.forename: David W
authors.initials: DW
authors.lastname: Scott
authors.forename: Taylor S
authors.initials: TS
authors.lastname: Dunn
authors.forename: Mary E
authors.initials: ME
authors.lastname: Ballestas
authors.forename: Silvio H
authors.initials: SH
authors.lastname: Litovsky
authors.forename: Rakesh P
authors.initials: RP
authors.lastname: Patel
mesh_terms: D000818:Animals; D019556:COS Cells; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D002522:Chlorocebus aethiops; D003599:Cytoskeleton / Q000502:physiology; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D006031:Glycosylation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009000:Monocytes / Q000502:physiology*; D020033:Protein Isoforms; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D020033:Protein Isoforms; D018799:Intercellular Adhesion Molecule-1
keywords: N-glycan; adhesion molecule; endothelial cell; inflammation