| Project/Area Number |
16570117
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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 |
Functional biochemistry
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| Research Institution | Kagawa University |
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Principal Investigator |
NISHI Nozomu Kagawa University, Faculty of Medicine, Department of Endocrinology, Research Associate, 医学部, 助手 (10145047)
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| Co-Investigator(Kenkyū-buntansha) |
東海林 博樹 香川大学, 医学部, 助手 (10263873)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | galectin / matrix metalloproteinase / oligosaccharide / lectin / protease / glycoprotein / neutrophil / inflammation / oligoaccharide |
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| Research Abstract |
1.Oligosaccharide structures of pro-MMP-9 recognized by galectin-8 : It has been reported that pro-MMP-9 has three N-linked oligosaccharides and many O-linked oligosaccharides. We have tested the effect of removal of the N-linked oligosaccharides on the interaction with galectin-8.pro-MMP-9 deficient in N-linked oligosaccharides retained the ability to interact with galectin-8 suggesting that O-linked oligosaccharides are important in galectin-8-mediated activation of pro-MMP-9. However, removal of O-linked oligosaccharides of pro-MMP-9 with a combination of O-glycanase, sialidase A and β(1-4)galactosidase did not abolish the ability to interact with galectin-8. These results suggest that galectin-8 can bind to both N- and O-linked oligosaccharides of proMMP-9.
2.Mechanism for acceleration of pro-MMP-9 activation by galectin-8 : We have already reported that N-terminal carbohydrate recognition domain (N-CRD) of galectin-8 has high affinity for pro-MMP-9 and that inactivation of either N- or C-CRD by site-directed mutagenesis resulted in loss of the activity (acceleration of pro-MMP-9 activation by MMP-3). MMP-3 binds to both N- and C-CRD of galectin-8 with similar affinities. In addition, we found that tag-free galectin-8 exhibits lower activity than that of GST-galectin-8. Based on these results, we speculate that galectin-8 increases the probability of interaction between pro-MMP-9 and MMP-3 in dilute solutions by binding both components (pro-MMP-9 at N-CRD and MMP-3 at C-CRD). The effect of GST-moiety can be explained by stable dimer formation of GST-galectin-8 but not tag-free galectin-8, which further increases the probability of the interaction.
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