通过对由分离的微绒毛核心成核的肌动蛋白丝进行电子显微镜观察，直接测量肌动蛋白聚合速率常数。
Direct measurement of actin polymerization rate constants by electron microscopy of actin filaments nucleated by isolated microvillus cores.
作者信息
Pollard T D, Mooseker M S
出版信息
J Cell Biol. 1981 Mar;88(3):654-9. doi: 10.1083/jcb.88.3.654.
Abstract
We used actin filament bundles isolated from intestinal brush-border microvilli to nucleate the polymerization of pure muscle actin monomers into filaments. Growth rates were determined by electron microscopy by measuring the change in the length of the filaments as a function of time. The linear dependence of the growth rates on the actin monomer concentration provided the rate constants for monomer association and dissociation at the two ends of the growing filament. The rapidly growing ("barbed") end has higher association and dissociation rate constants than the slowly growing ("pointed") end. The values of these rate constants differ in 20 mM KCl compared with 75 mM KCl, 5 mM MgSO4. 2 microM cytochalasin B blocks growth entirely at the barbed end, apparently by reducing both association and dissociation rate constants to near zero, but inhibits growth at the pointed end to only a small extent.
摘要
我们使用从肠道刷状缘微绒毛分离出的肌动蛋白丝束，使纯肌肉肌动蛋白单体聚合成丝。通过电子显微镜测量丝长度随时间的变化来确定生长速率。生长速率对肌动蛋白单体浓度的线性依赖性提供了生长丝两端单体结合和解离的速率常数。快速生长的（“带刺的”）末端比缓慢生长的（“尖锐的”）末端具有更高的结合和解离速率常数。与75 mM KCl、5 mM MgSO4相比，在20 mM KCl中这些速率常数的值有所不同。2 microM细胞松弛素B完全阻止带刺末端的生长，显然是通过将结合和解离速率常数都降低到接近零，但仅在很小程度上抑制尖锐末端的生长。
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