FLUORESCENT ACTIN-FILAMENTS MOVE ON MYOSIN FIXED TO A GLASS-SURFACE
FLUORESCENT ACTIN-FILAMENTS MOVE ON MYOSIN FIXED TO A GLASS-SURFACE
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DOI：
10.1073/pnas.83.17.6272
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发表时间：
1986-09-01
影响因子：
11.1
通讯作者：
SPUDICH, JA
中科院分区：
文献类型：
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作者：
KRON, SJ;SPUDICH, JA
用荧光鬼笔环肽稳定的单个肌动蛋白丝在固定于表面的肌球蛋白丝上表现出ATP依赖性运动。在pH 7.4和24 ℃下，C时，骨骼肌肌球蛋白的运动速率平均为3-4 μ m/s，网骨藻肌球蛋白的运动速率平均为1-2 μ m/s。这些速率与我们以前的肌球蛋白运动测定中测量的速率非常相似。运动的速率与所用肌动蛋白的类型相对无关。细丝速度显示在7.0和9.0之间的宽pH最适值，并且半最大速度所需的ATP浓度为50 μ M。有证据表明，肌动蛋白在肌球蛋白上的运动最多需要单个粗丝中的头数。该系统提供了一个实用的，定量肌球蛋白运动与纯化的蛋白质测定。
Single actin filaments stabilized with fluorescent phalloidin exhibit ATP-dependent movement on myosin filaments fixed to a surface. At pH 7.4 and 24.degree. C, the rates of movement average 3-4 .mu.m/s with skeletal muscle myosin and 1-2 .mu.m/s with Dictyostelium myosin. These rates are very similar to those measured in our previous myosin movement assays. The rates of movement are relatively independent of the type of actin used. The filament velocity shows a broad pH optimum between 7.0 and 9.0, and the concentration of ATP required for half-maximal velocity is 50 .mu.M. Evidence was obtained to suggest that movement of actin over myosin requires at most the number of heads in a single thick filament. This system provides a practical, quantitative myosin-movement assay with purified proteins.