Matrix-assisted laser desorption/ionization mass spectrometry compatible beta-elimination of O-linked oligosaccharides.
A new beta-elimination procedure has been introduced to cleave O-linked oligosaccharides from low- to sub-microgram amounts of glycoproteins prior to analysis by mass spectrometry. Borane-ammonia complex in aqueous ammonia is used as a cleaving solution alternative to the sodium borohydride/sodium hydroxide medium conventionally used in beta-elimination. The procedure results in minimum sample purification, leading to minimal sample loss and consequently an overall enhancement in sensitivity. It was applied successfully in the analysis of bovine fetuin and submaxillary mucin, as well as to a complex bile-salt-stimulated lipase glycoprotein isolated from human milk.
Huang
Yunping
Y
Department of Chemistry, Indiana University, Bloomington, IN 47405, USA.
Konse
Tomonori
T
Mechref
Yehia
Y
Novotny
Milos V
MV
D000641:Ammonia; D000818:Animals; D001880:Boranes; D002417:Cattle; D003911:Dextrans / Q000737:chemistry; D005260:Female; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D008894:Milk Proteins / Q000032:analysis*; D008895:Milk, Human / Q000201:enzymology*; D009077:Mucins / Q000737:chemistry*; D009844:Oligosaccharides / Q000032:analysis*; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D002787:Sterol Esterase / Q000032:analysis*; D000509:alpha-Fetoproteins / Q000737:chemistry*
D001880:Boranes; D003911:Dextrans; D006023:Glycoproteins; D008894:Milk Proteins; D009077:Mucins; D009844:Oligosaccharides; D011134:Polysaccharides; D000509:alpha-Fetoproteins; D000641:Ammonia; C054784:bile salt-stimulated lipase; D002787:Sterol Esterase