TAILIEUCHUNG - Báo cáo khóa học: Differential carbohydrate epitope recognition of globotriaosyl ceramide by verotoxins and a monoclonal antibody

The role of renal expression of the glycosphingolipid verotoxin receptor, globotriaosylceramide, in susceptibility to verotoxin-induced hemolytic uremic syndrome is unclear. We show that a single glycosphingolipid can discriminate multiple specific ligands. Antibody detection of globo-triaosylceramide in renal sections does not necessarily pre-dict verotoxin binding. | Eur. J. Biochem. 271 405-417 2004 FEBS 2004 doi Differential carbohydrate epitope recognition of globotriaosyl ceramide by verotoxins and a monoclonal antibody Role in human renal glomerular binding Davin Chark1 2 Anita Nutikka1 Natasha Trusevych1 3 Julia Kuzmina1 3 and Clifford Lingwood1 2 3 1 Research Institute Division of Infection Immunity Injury and Repair The Hospital for Sick Children Ontario Canada department of Laboratory Medicine Pathobiology and 3Department of Biochemistry University of Toronto Canada The role of renal expression of the glycosphingolipid verotoxin receptor globotriaosylceramide in susceptibility to verotoxin-induced hemolytic uremic syndrome is unclear. We show that a single glycosphingolipid can discriminate multiple specific ligands. Antibody detection of globo-triaosylceramide in renal sections does not necessarily predict verotoxin binding. The deoxyglobotriaosylceramide binding profile for verotoxin 1 verotoxin 2 and monoclonal anti-globotriaosylceramide are distinct. Anti-globotriaosyl-ceramide had greater dependency on the intact a-galactose and reducing glucose of globotriaosylceramide than vero-toxin 1 while verotoxin 2 was intermediate. These ligands differentially stained human kidney sections. Glomerulopathy is the primary verotoxin-associated pathology in hemolytic uremic syndrome. For most samples verotoxin 1 immunostaining within adult glomeruli was observed type A . Some samples however lacked glomerular binding type B . Anti-globotriaosylceramide and less effectively verotoxin 2 stained all glomeruli. Verotoxin 1 anti-globo-triaosylceramide tubular staining was comparable. Type B glomerular tubular globotriaosylceramide showed minor but significant fatty acid compositional differences. Vero-toxin 1 type B glomerular binding became evident following pretreatment with cold acetone or methyl-b-cyclodextrin used to deplete cholesterol. Direct visualization using fluorescein .

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