大肠杆菌 MreB 蛋白的纯化与表征。
Purification and characterization of Escherichia coli MreB protein.
机构信息
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA.
出版信息
J Biol Chem. 2013 Feb 1;288(5):3469-75. doi: 10.1074/jbc.M112.413708. Epub 2012 Dec 12.
Abstract
The actin homolog MreB is required in rod-shaped bacteria for maintenance of cell shape and is intimately connected to the holoenzyme that synthesizes the peptidoglycan layer. The protein has been reported variously to exist in helical loops under the cell surface, to rotate, and to move in patches in both directions around the cell surface. Studies of the Escherichia coli protein in vitro have been hampered by its tendency to aggregate. Here we report the purification and characterization of native E. coli MreB. The protein requires ATP hydrolysis for polymerization, forms bundles with a left-hand twist that can be as long as 4 μm, forms sheets in the presence of calcium, and has a critical concentration for polymerization of 1.5 μM.
摘要
肌动蛋白同源物 MreB 在杆状细菌中对于维持细胞形状是必需的，并且与合成肽聚糖层的全酶密切相关。该蛋白据报道以各种形式存在于细胞表面下的螺旋环中，发生旋转，并在细胞表面的两个方向上以斑块形式移动。对大肠杆菌蛋白的体外研究受到其聚集倾向的阻碍。在这里，我们报告了天然大肠杆菌 MreB 的纯化和表征。该蛋白的聚合需要 ATP 水解，形成左旋扭曲的束，长度可达 4μm，在钙离子存在下形成薄片，并具有聚合的临界浓度为 1.5μM。
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2
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PLoS One. 2011;6(10):e25129. doi: 10.1371/journal.pone.0025129. Epub 2011 Oct 5.
3
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EMBO J. 2011 Sep 30;30(24):4931-41. doi: 10.1038/emboj.2011.358.
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