CHARACTERIZATION OF RECOMBINANT MURINE INTERLEUKIN-5 EXPRESSED IN CHINESE-HAMSTER OVARY CELLS

被引：3
作者：
KODAMA, S
ENDO, T
TSUJIMOTO, M
KOBATA, A
机构：
[1] UNIV TOKYO,INST MED SCI,DEPT BIOCHEM,4-6-1 SHIROKANEDAI,MINATO KU,TOKYO 108,JAPAN
[2] SUNTORY INST BIOMED RES,SHIMAMOTO CHO,OSAKA 618,JAPAN
来源：
GLYCOBIOLOGY | 1992年 / 2卷 / 05期
关键词：
INTERLEUKIN; 5; N-LINKED SUGAR CHAIN; RECOMBINANT GLYCOPROTEIN;
D O I：
10.1093/glycob/2.5.419
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We have purified recombinant murine interleukin 5 (rmIL-5) from the supernatant of Chinese hamster ovary cells. Each peptide fragment of the purified rmIL-5 generated by Achromobacter protease I digestion was characterized and glycosylation sites were determined. Although rmIL-5 contains three potential sites of N-linked glycosylation (Asn-26, Asn-55 and Asn-69), Asn-69 is not glycosylated. The oligosaccharides released from the protein by hydrazinolysis were fractionated by paper electrophoresis, lectin column chromatography and gel permeation chromatography, and their structures were analysed by sequential exoglycosidase digestion in combination with methylation analysis. The results indicated that they are a mixture of bi-, tri- and tetraantennary complex-type sugar chains with and without a fucose at the C-6 position of the proximal N-acetylglucosamine residue and high-mannose-type sugar chains. Although > 80% of the sugar chains are neutral oligosaccharides similar to recombinant human IL-5 (rhIL-5; Kodama,S., Endo,T., Tsuruoka,N., Tsujimoto,M. and Kobata,A. (1991) J. Biochem., 110, 693-701), rmIL-5 has more tetraantennary oligosaccharides than rhIL-5. A site differential study revealed that Asn-55 has more tetraantennary oligosaccharides than Asn-26.
引用
收藏
页码：419 / 427
页数：9
相关论文
共 31 条
  • [1] CLONING OF CDNA FOR HUMAN T-CELL REPLACING FACTOR (INTERLEUKIN-5) AND COMPARISON WITH THE MURINE HOMOLOG
    AZUMA, C
    TANABE, T
    KONISHI, M
    KINASHI, T
    NOMA, T
    MATSUDA, F
    YAOITA, Y
    TAKATSU, K
    HAMMARSTROM, L
    SMITH, CIE
    SEVERINSON, E
    HONJO, T
    [J]. NUCLEIC ACIDS RESEARCH, 1986, 14 (22) : 9149 - 9158
  • [2] CUMMING D A, 1991, Glycobiology, V1, P115, DOI 10.1093/glycob/1.2.115
  • [3] STRUCTURAL STUDY ON THE CARBOHYDRATE MOIETY OF HUMAN PLACENTAL ALKALINE-PHOSPHATASE
    ENDO, T
    OHBAYASHI, H
    HAYASHI, Y
    IKEHARA, Y
    KOCHIBE, N
    KOBATA, A
    [J]. JOURNAL OF BIOCHEMISTRY, 1988, 103 (01) : 182 - 187
  • [4] PRIMARY STRUCTURE OF A FUCOSE-SPECIFIC LECTIN OBTAINED FROM A MUSHROOM, ALEURIA-AURANTIA
    FUKUMORI, F
    TAKEUCHI, N
    HAGIWARA, T
    OHBAYASHI, H
    ENDO, T
    KOCHIBE, N
    NAGATA, Y
    KOBATA, A
    [J]. JOURNAL OF BIOCHEMISTRY, 1990, 107 (02) : 190 - 196
  • [5] GLASGOW LR, 1977, J BIOL CHEM, V252, P8615
  • [6] SYNTHESIS AND PROCESSING OF ASPARAGINE-LINKED OLIGOSACCHARIDES
    HUBBARD, SC
    IVATT, RJ
    [J]. ANNUAL REVIEW OF BIOCHEMISTRY, 1981, 50 : 555 - 583
  • [7] KAGAWA Y, 1988, J BIOL CHEM, V263, P17508
  • [8] CLONING OF COMPLEMENTARY-DNA ENCODING T-CELL REPLACING FACTOR AND IDENTITY WITH B-CELL GROWTH FACTOR-II
    KINASHI, T
    HARADA, N
    SEVERINSON, E
    TANABE, T
    SIDERAS, P
    KONISHI, M
    AZUMA, C
    TOMINAGA, A
    BERGSTEDTLINDQVIST, S
    TAKAHASHI, M
    MATSUDA, F
    YAOITA, Y
    TAKATSU, K
    HONJO, T
    [J]. NATURE, 1986, 324 (6092) : 70 - 73
  • [9] KOBATA A, 1972, METHOD ENZYMOL, V28, P262
  • [10] CARBOHYDRATE STRUCTURES OF HUMAN INTERLEUKIN-5 EXPRESSED IN CHINESE-HAMSTER OVARY CELLS
    KODAMA, S
    ENDO, T
    TSURUOKA, N
    TSUJIMOTO, M
    KOBATA, A
    [J]. JOURNAL OF BIOCHEMISTRY, 1991, 110 (05) : 693 - 701
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