| Project/Area Number |
13670114
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | Nagoya University |
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Principal Investigator |
MURAMATSU Hisako Nagoya University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 講師 (50182134)
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| Co-Investigator(Kenkyū-buntansha) |
MURAMATSU Takashi Nagoya University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (00030891)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | midkine / chondroitinsulfate / LRP / LDL interein receptor family / LRP6 / oversulfation / integrin / プロテインシークエンス / シャペロン |
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| Research Abstract |
FLAG-tagged LRP6, which belongs to the LDL receptor family, bound to midkine (MK) column with calcium dependent mannor as strongly as LRP. LRP and LRP6 are different in the intracellular domain, and the two receptors may utilize different signaling systems to transduce the signal derived from MK. Chondroitin 4-sulfate 6-sulfotransferase was produced in the Baculovirus system, and the recombinant enzyme was used to produce artificial E structure, which has 4,6-disulfated N-acetylgalactosamine. This artificial E structure was found to bind to MK strongly, indicating that other modifications are not required for strong binding to MK. MK binding proteins were isolated from Day 13 mouse embryos, and were identified by protein microsequencing after separation by SDS-PAGE. Consequently, the 110kDa binding protein was identified as β1-integrin. Further studies revealed that α4β1 and α6β1 integrin were present in the receptor complex for MK and that they bound directly to MK. Together with previous results, we conclude that the receptor complex for MK contains LDL receptor family members, proteoglycans such as PTPζ with chondroitin sutfate E structure and integrins.
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