Biological activity and structural stability of N-deglycosylated recombinant human erythropoietin.
Controversy exists regarding the functional role of N-linked oligosaccharides in the hormone erythropoietin. We have now examined the role of carbohydrates in the hormone's action using quantitative enzymatic deglycosylation. N-deglycosylated hormone exhibited full biological activity and potency in vitro. Denaturing with 6M urea and renaturing revealed that both the native and N-deglycosylated forms recovered full activity as long as the intrachain disulfide bonds remained intact. Therefore, receptor recognition, subsequent biological activity and maintenance of tertiary structure are intrinsic properties of the polypeptide chain of erythropoietin.
Sytkowski
A J
AJ
Department of Medicine, New England Deaconess Hospital, Harvard Medical School, Boston, Massachusetts 02215.
Feldman
L
L
Zurbuch
D J
DJ
D000818:Animals; D006224:Cricetinae; D004921:Erythropoietin / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D011487:Protein Conformation; D011489:Protein Denaturation; D011956:Receptors, Cell Surface / Q000378:metabolism; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism
D011956:Receptors, Cell Surface; D017467:Receptors, Erythropoietin; D011994:Recombinant Proteins; D004921:Erythropoietin