Knäuper Vera, Murphy Gillian
Dental School, Cardiff University, Cardiff, UK.
Methods Mol Biol. 2010;622:233-43. doi: 10.1007/978-1-60327-299-5_14.
The degradation of the extracellular matrix during development and in disease is thought to result from the combined action of several proteolytic enzyme systems, including the matrix metalloproteinases (MMPs), serine proteinases, and cysteine proteinases. The majority of the soluble MMPs are synthesized as proenzymes which require extracellular activation in order to gain proteolytic activity and the analysis of their activation mechanism is a prerequisite for understanding MMP-mediated proteolysis.The emphasis of this chapter is the provision of the experimental tools to study MMP activation in vitro and in cellular model systems. Hence, we use the activation of procollagenase-3 (proMMP-13) and progelatinase A (proMMP-2) as examples of the methods used.
在发育过程和疾病状态下,细胞外基质的降解被认为是由多种蛋白水解酶系统共同作用的结果,这些系统包括基质金属蛋白酶(MMPs)、丝氨酸蛋白酶和半胱氨酸蛋白酶。大多数可溶性MMPs以酶原形式合成,需要细胞外激活才能获得蛋白水解活性,对其激活机制的分析是理解MMP介导的蛋白水解作用的先决条件。本章重点是提供用于在体外和细胞模型系统中研究MMP激活的实验工具。因此,我们以原胶原酶-3(proMMP-13)和原明胶酶A(proMMP-2)的激活为例来说明所使用的方法。