Karlsson R
MRC Laboratory of Molecular Biology, Cambridge, U.K.
Gene. 1988 Sep 7;68(2):249-57. doi: 10.1016/0378-1119(88)90027-3.
Actin interacts with a number of so-called actin-binding proteins which participate at various stages of the cell motility process such as regulation of filament formation, assembly and disassembly of filaments, force generation and depolymerization. Gene technology makes a precise mapping of the interacting surfaces on the actin molecules possible by studying specifically designed actin mutants expressed in a suitable organism. In addition, the production of engineered actin will become increasingly important when the three-dimensional structure of actin is determined. Chicken beta-actin can be produced in large quantities in Escherichia coli but such actin shows only a limited biological activity and thus seems to be of minor interest in future studies of structure-function relationships of this molecule. To circumvent the problem of a denatured bacterial protein, the yeast Saccharomyces cerevisiae was chosen as an alternative organism to express actin. This paper describes the expression, isolation and characterization of the yeast-produced chicken beta-actin. From a 12-liter culture of yeast cells, 500 micrograms of polymerizable beta-actin was isolated.
肌动蛋白与许多所谓的肌动蛋白结合蛋白相互作用，这些蛋白参与细胞运动过程的各个阶段，如细丝形成的调节、细丝的组装和解聚、力的产生以及解聚作用。基因技术通过研究在合适生物体中表达的特定设计的肌动蛋白突变体，使得精确绘制肌动蛋白分子上的相互作用表面成为可能。此外，当确定肌动蛋白的三维结构时，工程化肌动蛋白的生产将变得越来越重要。鸡β-肌动蛋白可以在大肠杆菌中大量生产，但这种肌动蛋白仅表现出有限的生物学活性，因此在该分子结构-功能关系的未来研究中似乎不太受关注。为了解决细菌蛋白变性的问题，选择酿酒酵母作为表达肌动蛋白的替代生物体。本文描述了酵母产生的鸡β-肌动蛋白的表达、分离和表征。从12升酵母细胞培养物中分离出了500微克可聚合的β-肌动蛋白。
