Nie Jing, Pei Duanqing
Hope Heart Program, Benaroya Research Institute at Virginia Mason, Seattle, WA, USA.
Methods Mol Biol. 2010;622:99-110. doi: 10.1007/978-1-60327-299-5_6.
Matrix metalloproteinases (MMPs) modulate cell-cell and cell-matrix interactions and the membrane-type MMPs (MT-MMPs) are especially important for pericellular proteolysis. Understanding the activity, regulation, and function of MT-MMPs would provide important insight for ongoing research in many diseases including cancer, fibrosis, and autoimmune diseases. In this chapter, we introduce the methods to express and purify recombinant MT-MMPs using Escherichia coli or mammalian cells, as well as how to detect the activity of MT-MMPs in proMMP-2 activation using gelatin zymography.
基质金属蛋白酶(MMPs)可调节细胞间和细胞与基质间的相互作用,而膜型基质金属蛋白酶(MT-MMPs)对细胞周围蛋白水解尤为重要。了解MT-MMPs的活性、调节机制和功能,将为包括癌症、纤维化和自身免疫性疾病在内的多种疾病的现有研究提供重要见解。在本章中,我们介绍了使用大肠杆菌或哺乳动物细胞表达和纯化重组MT-MMPs的方法,以及如何利用明胶酶谱法检测MT-MMPs在激活前MMP-2中的活性。