Tait J F, Frieden C
Biochemistry. 1982 Nov 23;21(24):6046-53. doi: 10.1021/bi00267a004.
We examined the properties of rabbit skeletal muscle actin labeled at Cys-373 with N-ethylmaleimide or with (iodoacetamido)tetramethylrhodamine, and of N-ethylmaleimide-actin further modified with 7-chloro-4-nitro-2,1,3-benzoxadiazole (which primarily labels Lys-372). All three derivatives polymerize more rapidly than unlabeled actin. As measured by fluorescence photobleaching recovery and low-shear viscometry, all three also show a lower extent of network formation relative to native actin. N-Ethylmaleimide has a much smaller effect on the rate of polymerization and on network formation than do the other two derivatives. We suggest that chemical modification of actin with these compounds may stabilize nuclei, accounting for the acceleration of polymerization. Stabilization of nuclei also reduces the average filament length at equilibrium, thereby reducing the extent of network formation. We note a parallel between these results and the effects that cytochalasin and capping proteins have on the polymerization of actin.
我们研究了用N-乙基马来酰亚胺或(碘乙酰胺基)四甲基罗丹明标记于半胱氨酸-373位点的兔骨骼肌肌动蛋白,以及用7-氯-4-硝基-2,1,3-苯并恶二唑(主要标记赖氨酸-372)对N-乙基马来酰亚胺-肌动蛋白进行进一步修饰后的产物的性质。这三种衍生物的聚合速度均比未标记的肌动蛋白快。通过荧光漂白恢复和低剪切粘度测定法测量,相对于天然肌动蛋白,这三种衍生物形成网络的程度也较低。与其他两种衍生物相比,N-乙基马来酰亚胺对聚合速率和网络形成的影响要小得多。我们认为,用这些化合物对肌动蛋白进行化学修饰可能会稳定核,这可以解释聚合加速的现象。核的稳定也会降低平衡时的平均细丝长度,从而减少网络形成的程度。我们注意到这些结果与细胞松弛素和封端蛋白对肌动蛋白聚合的影响之间存在相似之处。