SUMO蛋白轉譯後修飾之基質金屬蛋白酶-7在細胞核中參與核纖層蛋白蛋白質裂解之研究

The nucleus translocation of SUMOylated MMP-7 mediated proteolysis of nucleus matrix protein lamin B1

蔡寧馨(Ning-Xing Tsai)

指導教授 ： 游偉絢

國立臺灣大學/醫學院/生物化學暨分子生物學研究所/碩士(2010年)

https://doi.org/10.6342/NTU.2010.10407

摘要

無資料

關鍵字

基質金屬蛋白酶-7 ； 細胞核基質 ； SUMO修飾 ； 核纖層蛋白B1 ； 細胞癌化

並列摘要

Matrix metalloptroteinase-7 (MMP-7; matrilysin) is a member of the MMPs family, which is involved in the extracellular matrix (ECM) component degradation and tissue remodeling. MMP-7 was reported as a secreted enzyme localized in the epical surface of normal glandular epithelial cells by Western blot and zymography analysis. Interestingly, a 31 kDa protein with protease activity found in nucleus was identified as pro-MMP-7. Furthermore, the protein sequence of MMP-7 was analyzed and a putative sumoylation site was defined by Dr. Wei-Hsuan Yu. (1998 Gordon Research Conferences poster)
To determine the sumoylation state of MMP-7, we used two anti-MMP7 antibodies to immunoprecipitate MMP-7, and then analyzed by Western blot with an anti-SUMO3 antibody. These results demonstrated that MMP-7 was modified by SUMO3. The prominent SUMO3 modified MMP-7 in the nucleus fraction implicated that SUMOylation post-translational modification for MMP-7 could be a nucleus trafficking signature for MMP-7. Also, by analyzing the total cell lysate with immunoprecipitation method, we found that MMP-7 were also ubiquitinated. The SUMO3 and ubiquitin tagged on MMP-7 could be responsible for MMP-7 intracellular trafficking route. The detail mechanism remains for further investigation.
After the nuclear translocation of MMP-7 was verified, we further found that lamin B1 became fragmented when MMP-7 was overexpressed. Using an anti-lamin B1 antibody, the immunoprecipitation data showed that the active site mutated MMP-7 was pulled down and was detected by Western blotting. Furthermore, the intact structure of lamin B1 at nuclear lamina was changed when MMP-7 was overexpressed. My data suggested that lamin B1 could be a substrate of MMP-7 in the nucleus. As the MMP-7/SUMO3 overexpressed cells formed colonies in the soft agar assay, the SUMOylated MMP-7 might involved in cancer progression by chromatin remodeling. The detailed underlying mechanism will be addressed in the near future.

並列關鍵字

MMP-7 ； SUMOylation ； laminB1 ； nuclear matrix ； tumor progression

參考文獻

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