Blood group A glycolipid antigen biosynthesis: discrimination between biosynthesized and enzyme preparation derived blood group A antigen by mass spectrometry.
A monofucosyl type 1 chain blood group A hexaglycosylceramide was biosynthesized in solution using the type 1 chain blood group H pentaglycosylceramide as precursor, a crude microsomal fraction prepared from the mucosa scraping of a blood group A pig small intestine as enzyme source, and uridine diphosphate-N-acetyl-(1-14C)galactosamine as sugar donor. The radioactive product was enriched using reversed-phase column chromatography and silica gel HPLC. The peak, as detected by a beta-flow scintillation counter, was collected, permethylated, and analyzed by mass spectrometry. Carbohydrate sequence ions were found, indicating the presence of both the biosynthesized and a native, non-14C-containing blood group A hexaglycosylceramide. The blood group A pig small intestinal mucosa used as the enzyme source contain blood group A hexaglycosylceramide as the predominant glycolipid. Therefore, it is concluded that the nonradioactive blood group A hexaglycosylceramide found after the biosynthesis is derived from the enzyme preparation.
Holgersson
J
J
Department of Medical Biochemistry, University of Göteborg, Sweden.
Jacobsson
A
A
Breimer
M E
ME
Samuelsson
B E
BE
D000017:ABO Blood-Group System; D000818:Animals; D001345:Autoradiography; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D006017:Glycolipids / Q000302:isolation & purification; D013058:Mass Spectrometry; D008745:Methylation; D058428:Silica Gel; D012822:Silicon Dioxide; D013552:Swine
D000017:ABO Blood-Group System; D006017:Glycolipids; D058428:Silica Gel; D012822:Silicon Dioxide