Role of glycosylation on the secretion and biological activity of erythropoietin.
Biochemistry
; 31(41): 9871-6, 1992 Oct 20.
Article
em En
| MEDLINE
| ID: mdl-1390770
ABSTRACT
The erythropoietin (EPO) molecule contains four carbohydrate chains. Three contain N-linkages to asparagines at positions 24, 38, and 83, and one contains an O-linkage to a serine at position 126. We constructed human EPO variants that eliminated the three N-glycosylation sites by replacing the asparagines with glutamines singly or in combination. The O-linked carbohydrate chain was removed by replacing the serine with glutamine, valine, histidine, or alanine. A variant with a double mutation (Gln38,83) and another with a triple mutation (Gln24,38,83) were secreted poorly from COS1 and CHO cells even though RNA encoding these variants was present. All other variants with mutations in N-linked glycosylation sites were secreted normally. Removal of any of the N-glycosylation sites reduced the in vivo but not the in vitro biological activity of the EPO molecule. All the mutations at Ser126, the O-glycosylation site, were secreted normally. In vitro activity was also unaffected except for Ala126 which had a 50-fold decrease. The Val126 variant was tested in vivo, and its specific activity was only slightly less than that of the native EPO, which indicates that the O-linked carbohydrate is not essential for activity.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Carboidratos
/
Eritropoetina
Limite:
Animals
Idioma:
En
Revista:
Biochemistry
Ano de publicação:
1992
Tipo de documento:
Article