Birkedal-Hansen Henning, Yamada Susan, Windsor Jack, Poulsen Anne Havemose, Lyons Guy, Stetler-Stevenson William, Birkedal-Hansen Bente
National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland, USA.
Curr Protoc Cell Biol. 2003 Feb;Chapter 10:Unit 10.8. doi: 10.1002/0471143030.cb1008s17.
Matrix metalloproteinases are a class of enzymes that play an important role in the remodeling of the extracellular matrix in development and cancer metastasis. This unit describes a set of methods-cell-mediated dissolution of type I collagen fibrils, direct and reverse zymography, enzyme capture based on a-2 macroglubulin and TIMP-1 and -2, and demonstration of crytic thiol groups in metalloproteinase precursors-that are used to characterize the functions of matrix metalloproteinases and their inhibitors.
基质金属蛋白酶是一类在发育和癌症转移过程中细胞外基质重塑中发挥重要作用的酶。本单元描述了一组方法——细胞介导的I型胶原纤维溶解、直接和反向酶谱分析、基于α-2巨球蛋白以及TIMP-1和TIMP-2的酶捕获,以及金属蛋白酶前体中半胱氨酸巯基的显示——这些方法用于表征基质金属蛋白酶及其抑制剂的功能。