Diaz-Valencia Juan Daniel, Bailey Megan, Ross Jennifer L
Department of Physics, University of Massachusetts Amherst, Amherst, Massachusetts, USA.
Methods Cell Biol. 2013;115:191-213. doi: 10.1016/B978-0-12-407757-7.00013-X.
Microtubule-severing enzymes are a novel class of microtubule regulators. They are enzymes from the ATPases associated with various cellular activities family (AAA+) that utilize ATP to cut microtubules into smaller filaments. Discovered over 20 years ago, there are still many open questions about severing enzymes. Both cellular and biochemical studies need to be pursued to fully understand how these enzymes function mechanistically in the cell. Here, we present methods to express, purify, and test the biophysical nature of these proteins in vitro to begin to address the biochemical and biophysical mechanisms of this important and novel group of microtubule destabilizers.
微管切断酶是一类新型的微管调节剂。它们是来自与各种细胞活动相关的ATP酶家族（AAA+）的酶，利用ATP将微管切割成更小的细丝。20多年前就已发现，但关于切断酶仍有许多未解决的问题。需要进行细胞和生化研究，以全面了解这些酶在细胞中的作用机制。在这里，我们介绍了在体外表达、纯化和测试这些蛋白质生物物理性质的方法，以开始探讨这一重要且新型的微管去稳定剂的生化和生物物理机制。
