裂殖酵母 Schizosaccharomyces pombe 肌动蛋白的纯化及其与肌动蛋白功能差异的表征。
Purification of actin from fission yeast Schizosaccharomyces pombe and characterization of functional differences from muscle actin.
机构信息
Department of Molecular Cellular and Developmental Biology, Yale University, New Haven, Connecticut 06520-8103, USA.
出版信息
J Biol Chem. 2011 Feb 18;286(7):5784-92. doi: 10.1074/jbc.M110.199794. Epub 2010 Dec 9.
Abstract
Fission yeast Schizosaccharomyces pombe is an important genetic model organism for studying the mechanisms of endocytosis and cytokinesis. However, most work on the biochemical properties of fission yeast actin-binding proteins has been done with skeletal muscle actin for matters of convenience. When simulations of mathematical models of the mechanism of endocytosis were compared with events in live cells, some of the reactions appeared to be much faster than observed in biochemical experiments with muscle actin. Here, we used gelsolin affinity chromatography to purify actin from fission yeast. S. pombe actin shares many properties with skeletal muscle actin but has higher intrinsic nucleotide exchange rate, faster trimer nucleus formation, faster phosphate dissociation rate from polymerized actin, and faster nucleation of actin filaments with Arp2/3 complex. These properties close the gap between the biochemistry and predictions made by mathematical models of endocytosis in S. pombe cells.
摘要
裂殖酵母 Schizosaccharomyces pombe 是研究胞吞作用和胞质分裂机制的重要遗传模式生物。然而，由于方便起见，大多数关于裂殖酵母肌动蛋白结合蛋白生化特性的研究都是使用骨骼肌肌动蛋白进行的。当内吞作用机制的数学模型模拟与活细胞中的事件进行比较时，一些反应似乎比用骨骼肌肌动蛋白进行的生化实验观察到的要快得多。在这里，我们使用凝胶电泳亲和层析从裂殖酵母中纯化肌动蛋白。裂殖酵母肌动蛋白与骨骼肌肌动蛋白有许多共同的特性，但具有更高的内在核苷酸交换率、更快的三聚体核形成、更快的聚合肌动蛋白从磷酸盐解离的速率以及更快的与 Arp2/3 复合物形成肌动蛋白丝的成核作用。这些特性缩小了裂殖酵母细胞内吞作用的生物化学和数学模型预测之间的差距。
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