| Project/Area Number |
09356010
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Applied molecular and cellular biology
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MORI Yuji (1998-1999) Graduate School of Agricultural and Life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (40157871)
小川 智也 (1997) 東京大学, 大学院・農学生命科学研究科, 教授 (30087572)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Tateki Natural Products Research Dept, Teikoku Hormone Mfg. Co. Ltd., Researcher, 研究本部・天然物研究部, 主任研究員
NUKATA Tomoo RIKEN the Institute of Physical and Chemical Research, Researcher, 細胞制御科学研究科, 専任研究員
SHIOTA Kunio Graduate School of Agricultural and Life Sciences, The University of Tokyo, Professor, 大学院・農学生命科学研究科, 教授 (80196352)
能田 健 鳥取大学, 農学部, 助教授 (60218287)
平田 統一 岩手大学, 農学部, 助手 (20241490)
森 裕司 東京大学, 大学院・農学生命科学研究科, 教授 (40157871)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥30,300,000 (Direct Cost: ¥30,300,000)
Fiscal Year 1999: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1998: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1997: ¥16,700,000 (Direct Cost: ¥16,700,000)
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| Keywords | equine chorionic gonadotropin (eCG) / equine follicle stimulating hormone (eFSH) / glycosylation / N-linked oligosaccharide / eCG / N-型複合型糖鎖 / 糖鎖合成法 / 変異遺伝体 / 活性評価 |
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| Research Abstract |
Equine chorionic gonadotropin (eCG) consists of highly glycosylated alpha-and beta-subunits and belongs to the member of the glycoprotein hormone family which includes luteinizing hormone (LH) and follicle-stimulating hormone (FSH). eCG is a unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in others species than the horse. The correct conformation of the heterodimer is also important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. As an approach to produce recombinant eCG with a potent FSH-like activity, we analyzed the roles of N-linked oligosaccharides in the biological activities of eCG and found that the FSH-like activity of eCG is resistant to the removal of N-linked oligosaccharides. We also investigated whether alpha and beta subunits can be synthesized as a single polypeptide chain (tethered-eCG) with any biological activities by using the tethered-eCG molecule in which the carboxyl terminus of the eCG beta-subunit was fused to the amino terminus of the alpha-subunit. The tethered-eCG was efficiently secreted from the transiently transfected CHO-K1 cells and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. Our data for the first time suggest that the tethered-eCG can be expressed efficiently in the CHO-K1 cells without losing LH-and FSH-like activities. Our results also suggest that the recombinant eCG protein with a potent FSH-like activity can possibly be pruduced by E. coli in which very little of the glycosylation occurs.
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