Analysis of the five glycosylation sites of human alpha 1-acid glycoprotein.
Orosomucoid (OMD) contains complex bi-, tri- and tetra-antennary glycan chains. Subfractionation of OMD into three molecular variants using concanavalin A lectin chromatography is based on variations in these complex structures. Standard h.p.l.c. profiles have been developed to analyse the percentage and distribution of the glycoforms present at each glycosylation site in OMD and its molecular variants. The ability to quantify the glycoforms present at each site allows us to extend the earlier results of others and resolve the remaining questions concerning the glycan structures of these variants. Most significantly, the proportions of bi-, tri- and tetra-antennary chains differ at each site for the three molecular variants. The most strongly retained variant from concanavalin A is uniquely capable of possessing biantennary chains at all five sites, whereas the unretained variant is completely devoid of biantennary chains. Only glycosylation site II of the five present is 100% biantennary in the retained and weakly retained variants. In addition, the two gene products of OMD were differentially glycosylated. Molecular masses of the glycoforms were verified by matrix-assisted u.v. laser desorption mass spectrometry. On the basis of the site distribution of oligosaccharides in the variants, efforts were made to understand the factors that control the processing of the carbohydrate chains in OMD. The results indicate that the 'site-directed' model of processing offers the most consistent explanation for the structures seen at the individual glycosylation sites of OMD.
Treuheit
M J
MJ
Department of Chemistry, University of Cincinnati, OH 45221-0172.
Costello
C E
CE
Halsall
H B
HB
D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A; D006031:Glycosylation; D006601:Hexoses / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009961:Orosomucoid / Q000378:metabolism / Q000648:ultrastructure*; D010455:Peptides / Q000302:isolation & purification; D011134:Polysaccharides; D012697:Serine Endopeptidases / Q000378:metabolism
D006601:Hexoses; D009961:Orosomucoid; D010455:Peptides; D011134:Polysaccharides; D003208:Concanavalin A; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase