Abstract
The asparagine-linked sugar chains of natural interferon-beta 1 secreted from human foreskin fibroblasts by poly I:poly C induction and of three recombinant human interferon-beta 1 produced by Chinese hamster ovary cells, mouse epithelial cells (C127), and human lung adenocarcinoma cells (PC8) were released quantitatively as oligosaccharides by hydrazinolysis followed by N-acetylation. After being reduced with either NaB3H4 or NaB2H4, their structures were comparatively analyzed. More than 80% of the sugar chains of natural interferon-beta 1 occur as biantennary complex-type sugar chains, approximately 10% of which contain N-acetyllactosamine repeating structure in their outer chain moieties. The remainders are 2,4- and 2,6-branched triantennary complex-type sugar chains. The sugar chains of the recombinant interferon-beta 1 derived from Chinese hamster ovary cells were very similar to those of its natural counterpart. In contrast, two other recombinant proteins contain quite different sugar chains. The protein derived from C127 cells contains complex-type sugar chains with the Gal alpha 1----3Gal beta 1----4GlcNAc group in their outer chain moieties. Their sialic acid residues occur solely as the Sia alpha 2----6Gal group, where Sia is sialic acid. In contrast, the sialic acid residues of other interferon-beta 1 occur as the Sia alpha 2----3Gal group only. A part of the sugar chains of the protein derived from PC8 cells contains bisecting N-acetylglucosamine residue in addition to the Gal alpha 1----3Gal beta 1----4GlcNAc group.
Highlights
The major parts of oligosaccharides from four samples were all recovered in the AAL' fraction; 91% for the natural HuIFN-Pl, and93%, 84%, and 78% for the CHO-, C127-, and PC8-rHuIFN-@lr,espectively
From the $Department of Biochemistry, the Institute of Medical Science,the University of Tokyo, Minuto-ku, Tokyo 108, Japan, $Toray Research Center Inc. and TBasic Research Laboratories, Toray Industries, Inc., 1111, Tebiro, Kamakura, Kanngawa 248, Japan, and the [[Departmentof Biology, Faculty of Education, Gunma University, Maebashi 371, Japan
Chinese hamster ovary cells have been used for moieties
Summary
The major parts of oligosaccharides from four samples were all recovered in the AAL' fraction; 91% for the natural HuIFN-Pl, and93%, 84%, and 78% for the CHO-, C127-, and PC8-rHuIFN-@lr,espectively. The lectin column fractions obtained from the fractions N from natural HuIFN-@1 anCdHO- and PC8-rHuIFN-Blgave almost the same elution patterns from Bio-Gel P-4 as fractions AN from respective samples. Elution patterns of the AALfractions obtained from natural HuIFN-@l and CHO- and C127-HuIFN-Pl from Bio-Gel P-4 column were very similar to those of the respective AAL+ fractions, except that the elution position of each peak was smaller by 1 glucose unit than the corresponding peak from AAL' fraction (data not shown).
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