1999 Fiscal Year Final Research Report Summary
Activation of matrix metalloproteinases on cell surface
| Project/Area Number |
10044244
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pathological medical chemistry
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SEIKI Motoharu Inst. Med. Sci., Univ. Tokyo, Prof., 医科学研究所, 教授 (10154634)
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| Co-Investigator(Kenkyū-buntansha) |
ITOH Yoshifumi Inst. Med. Sci., Univ. Tokyo, Assis. Prof., 医科学研究所, 助手 (70292852)
OKADA Akio Inst. Med. Sci., Univ. Tokyo, Assis. Prof., 医科学研究所, 助手 (00233320)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Extracellular matrix / Invasion and metastasis / Matrix metalloproteinase / Gelatinase A / MT-MMP |
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| Research Abstract |
(1) Malignant melanoma cells degrade gelatin gel beneath the cells by extending protrusions called invadpodia. We foud that MT1-MMP localizes on the invadpodia. To monitor the cell surface. MT1-MMP more easily, we made MT1MMP fused with GFP and expressed it in the melanoma cells. Localization of GFP/MT1-MMP was clearly observed at invade podia where actin bundle can be seen. MT1-MMP was also detected along with actin stress fibers when it is expressed in on invasive CHO-K1 cells. Thus, localization of MT1-nMMP seems to be regulated through actin cytoskelton within the cells.
(2) MT1-MMP activates pro-gelatinase Aby introducing cleavage in the propeptide domain in vitro. Expression of MT1-MMP in cultured cells induce cell-mediated activation of pro-gelatinase A. To confirm whether MT1-MMP can act as the activator of pro-gelatinase A in vivo, we generatedgene knockout mice by substituting the 1st to 4th exons of mouse genome to LacZ and Neo gene. Wild type mouse express MT1-MMP at high levels in various tissue during embryogenesis and activation of pro-gelatinase A in the tissue can be observed. In the knockout mice, however, such actibation of pro-gelatinase A could not be observed anymore. Thus, we conclued that MT1-MMP is the physiological activator of pro-gelatinase A.
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