Oligosaccharide mapping reveals hormone-specific glycosylation patterns on equine gonadotropin alpha-subunit Asn56
| Autor: | R. Russell Gotschall, George R. Bousfield |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Glycosylation
medicine.drug_class Molecular Sequence Data Alpha (ethology) Oligosaccharides Amidohydrolases chemistry.chemical_compound Endocrinology medicine Animals Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Glycoside hydrolase Amino Acid Sequence Cyanogen Bromide Disulfides Horses G alpha subunit chemistry.chemical_classification Edman degradation Oligosaccharide Molecular biology Enzyme chemistry Biochemistry Glycoprotein Hormones alpha Subunit Electrophoresis Polyacrylamide Gel Gonadotropin Asparagine |
| Zdroj: | Endocrinology. 137(6) |
| ISSN: | 0013-7227 |
| Popis: | Equine gonadotropin alpha-subunit glycosylation was examined by releasing oligosaccharides using a sequential enzymatic deglycosylation protocol and comparing the released oligosaccharide populations using a high resolution oligosaccharide mapping technique. Digestion of native alpha-subunit preparations with peptide-N-glycosidase altered their mobilities during SDS-PAGE under reducing conditions to positions intermediate between the corresponding native alpha-subunit and completely deglycosylated alpha-subunit bands. Complete alpha-subunit deglycosylation required reduction of disulfide bonds. Results of solid-phase Edman degradation demonstrated that partial deglycosylation occurred exclusively at Asn56. Oligosaccharide mapping of total oligosaccharides obtained by enzymatic deglycosylation of reduced, carboxymethylated alpha-subunit preparations revealed hormone-specific patterns of glycosylation in eLH alpha and eCG alpha. Oligosaccharide mapping of individual glycosylation sites revealed that hormone-specific glycosylation was primarily restricted to Asn56 of both subunit preparations and revealed a hormone-specific pattern of Asn56 glycosylation in eFSH alpha that was obscured in the total oligosaccharide map. eLH alpha Asn56 oligosaccharides appeared to be primarily seven variants of a monoantennary structure. eCG alpha Asn56 oligosaccharides consisted of one of two forms, either a sialylated biantennary oligosaccharide that appeared identical to a commercial carbohydrate standard or a lactosamine variant of that structure. |
| Databáze: | OpenAIRE |
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