2005 Fiscal Year Final Research Report Summary
Analysis of the ectodomain shedding mechanism of HB-EGF
| Project/Area Number |
16207014
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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 | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Osaka University |
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Principal Investigator |
MEKADA Eisuke Osaka University, Research Institute for Microbial Disease, Professor, 微生物病研究所, 教授 (20135742)
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| Co-Investigator(Kenkyū-buntansha) |
IWAMOTO Ryo Osaka University, Research Institute for Microbial Disease, Associate Professor, 微生物病研究所, 助教授 (10213323)
MIZUSHIMA Hiroto Osaka University, Research Institute for Microbial Disease, Research Associate, 微生物病研究所, 助手 (30379094)
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| Project Period (FY) |
2004 – 2005
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| Keywords | HB-EGF / ectodomain shedding / protease / ADAM |
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| Research Abstract |
Ectodomain shedding is critical process for the action of heparin-binding EGF-like growth factor (HB-EGF). This process is regulated by a number of extracellular stimuli and intracellular signaling molecules. However, the molecular mechanism for ectodomain shedding has largely remained to be clarified. We found the following results :
1)Over-expression of ADAM12 mutant form with defective metalloprotease domain induced ectodomain shedding of HB-EGF in Vero cells. Over-expression of several mutant forms of ADAM12 and co-precipitation experiments with anti-ADAM12 tag antibody suggested that ADAM12 may contribute to HB-EGF shedding as an adapter protein for signaling, rather than as a protease.
2)Screening of low molecular-weight chemicals which possess the inhibitory activity for HB-EGF ectodomain shedding was performed. A few compounds showed the inhibitory activity. Further characterization is underway.
3)To understand the molecules which are involved in ectodomain shedding of HB-EGF, we constructed a screening strategy which enables to identify genes necessary for the shedding. To do this, siRNA library containing 8.4 k human genome sequence was constructed and transfected in human cells by retro virus vecter system. Cell populations which did not respond to the stimuli for shedding were isolated.
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