| 肿瘤转移相关蛋白ST14的表达、纯化及活性鉴定 |
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| 引用本文： | 葛维挺,郑树,孙立峰,史影,胡涵光,丁克峰.肿瘤转移相关蛋白ST14的表达、纯化及活性鉴定[J].中国生物化学与分子生物学报,2004,20(5):685-689. |
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| 作者姓名： | 葛维挺 郑树 孙立峰 史影 胡涵光 丁克峰 |
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| 作者单位： | 1. 浙江大学生命科学学院,杭州,310029;浙江大学医学院附属二院肿瘤研究所,杭州,310009;杭州华大基因研发中心,杭州,310008
2. 浙江大学医学院附属二院肿瘤研究所,杭州,310009 |
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| 基金项目： | 国家自然科学基金资助项目 (No .3 0 2 0 0 3 2 5 )~~ |
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| 摘 要： | 细胞外基质包括基底膜和间隙基质 ,主要由胶原、糖蛋白和蛋白多糖等一些物质组成 ,具有维持细胞组织形态的作用 ,是细胞间相互作用的重要场所 .在肿瘤细胞的浸润和转移过程中 ,必须有细胞外基质的降解过程 ,研究发现多种蛋白酶与该过程有关 ,包括丝氨酸蛋白酶家族中的纤维蛋白溶酶原和纤维蛋白溶酶系统 ,金属蛋白酶系统中的MMP 2和MMP 9,组织蛋白酶B ,组织蛋白酶D ,以及透明质酸酶 ,胶原酶等1] .Ⅱ型跨膜丝氨酸蛋白酶 (TypeⅡtransmembraneserineprotease ,TTSP)ST14具有降解细胞外基质的能力 ,并能激活uPA前体及HGF SF前体 ,参与…
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| 收稿时间： | 2004-10-20 |
| 修稿时间： | 2003年9月30日 |
Expression and Purification of ST14, a Tumor Metastasis-associated Protein, and Its Activity Assay |
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| GE Wei-ting ,,ZHENG Shu ,SUN Li-feng ,SHI Ying ,HU Han-guang ,DING Ke-feng.Expression and Purification of ST14, a Tumor Metastasis-associated Protein, and Its Activity Assay[J].Chinese Journal of Biochemistry and Molecular Biology,2004,20(5):685-689. |
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| Authors: | GE Wei-ting ZHENG Shu SUN Li-feng SHI Ying HU Han-guang DING Ke-feng |
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| Institution: | ( 1) College of Life Sciences, Zhejiang University, Hangzhou 310029, China; 2) Cancer Institute, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009,China; 3) Hangzhou Genomics Institute, Hangzhou 310008,China |
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| Abstract: | ST14 is one of the type II transmembrane serine proteases that correlates with the process of tumor metastasis. The C-terminal catalytic region (900 bp) of ST14 was cloned into the expression vector pGEX-4T-2 and the positive plasmid pGEX-4T-2-ST14 was transformed into E. coli BL21, then cultured and induced with IPTG. The chaperonin GroEL was found to be tightly associated with the fusion protein and co-purified with it by regular GST affinity chromatography. A method for the removal of contaminating GroEL from GST-ST14 fusion protein was described, the purity of product was 96.2%. Enzyme activity assay indicated that this fusion protein had serine protease activity. |
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| Keywords: | ST14 gene fusion protein serine protease co-purification |
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