A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows.
A fluorescent resonant energy transfer-based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows.
复制标题
DOI：
10.1083/jcb.200110161
复制
发表时间：
2002-02-04
期刊：
影响因子：
--
通讯作者：
Chisholm RL
中科院分区：
文献类型：
--
作者：
Chew TL;Wolf WA;Gallagher PJ;Matsumura F;Chisholm RL
需要高空间和时间分辨率的方法来了解非肌肉肌球蛋白II在体内的调节。利用荧光共振能量转移，我们生产了一种新的生物传感器，可以同时测定活细胞中肌球蛋白轻链激酶（MLCK）的定位及其[Ca2+]4/钙调素结合状态。我们观察到弥漫性MLCK向应力纤维的瞬时招募及其在收缩前的原位激活。MLCK在迁移细胞的片层中高度活跃，而在收缩尾中则不活跃。这一意想不到的结果突出了mlck介导的肌球蛋白收缩在板层中作为迁移驱动力的潜在作用。在细胞质分裂过程中，MLCK在纺锤体赤道富集，在卵裂沟收缩前激活最多。当沟缩完成后，活跃的MLCK被重新分配到子细胞的两极。这些结果表明，MLCK是肌凝蛋白板层和收缩环的调节因子，并确定了肌凝蛋白功能可能由其他激酶介导的位点。
Approaches with high spatial and temporal resolution are required to understand the regulation of nonmuscle myosin II in vivo. Using fluorescence resonance energy transfer we have produced a novel biosensor allowing simultaneous determination of myosin light chain kinase (MLCK) localization and its [Ca2+]4/calmodulin-binding state in living cells. We observe transient recruitment of diffuse MLCK to stress fibers and its in situ activation before contraction. MLCK is highly active in the lamella of migrating cells, but not at the retracting tail. This unexpected result highlights a potential role for MLCK-mediated myosin contractility in the lamella as a driving force for migration. During cytokinesis, MLCK was enriched at the spindle equator during late metaphase, and was maximally activated just before cleavage furrow constriction. As furrow contraction was completed, active MLCK was redistributed to the poles of the daughter cells. These results show MLCK is a myosin regulator in the lamella and contractile ring, and pinpoints sites where myosin function may be mediated by other kinases.
登录
查看更多内容
影响因子：
4.8
作者：
Katoh, T;Morita, F
通讯作者：
Morita, F
影响因子：
4.8
作者：
Lin, PJ;Luby-Phelps, K;Stull, JT
通讯作者：
Stull, JT
影响因子：
4.8
作者：
Amano, M;Ito, M;Kaibuchi, K
通讯作者：
Kaibuchi, K
影响因子：
7.8
作者：
Beningo, KA;Dembo, M;Wang, YL
通讯作者：
Wang, YL
影响因子：
4.8
作者：
Lin, PJ;LubyPhelps, K;Stull, JT
通讯作者：
Stull, JT