嗜热真菌嗜热栖热放线菌驱动蛋白的克隆与表达。
Cloning and expression of kinesins from the thermophilic fungus Thermomyces lanuginosus.
作者信息
Sakowicz R, Farlow S, Goldstein L S
机构信息
Howard Hughes Medical Institute, Department of Cellular and Molecular Medicine, School of Medicine, University of California, San Diego, La Jolla 92093-0683, USA.
出版信息
Protein Sci. 1999 Dec;8(12):2705-10. doi: 10.1110/ps.8.12.2705.
The motor domain regions of three novel members of the kinesin superfamily TLKIF1, TLKIFC, and TLBIMC were identified in a thermophilic fungus Thermomyces lanuginosus. Based on sequence similarity, they were classified as members of the known kinesin families Unc104/KIF1, KAR3, and BIMC. TLKIF1 was subsequently expressed in Escherichia coli. The expression level was high, and the protein was mostly soluble, easy to purify, and enzymatically active. TLKIF1 is a monomeric kinesin motor, which in a gliding motility assay displays a robust plus-directed microtubule movement up to 2 microm/s. The discovery of TLKIF1 also demonstrates that a family of kinesin motors not previously found in fungi may in fact be used in this group of organisms.
在嗜热真菌嗜热栖热菌中鉴定出驱动蛋白超家族三个新成员TLKIF1、TLKIFC和TLBIMC的马达结构域区域。基于序列相似性，它们被归类为已知驱动蛋白家族Unc104/KIF1、KAR3和BIMC的成员。随后，TLKIF1在大肠杆菌中表达。表达水平很高，蛋白质大多可溶，易于纯化且具有酶活性。TLKIF1是一种单体驱动蛋白马达，在滑行运动测定中，它能以高达2微米/秒的速度沿着微管向正端强劲移动。TLKIF1的发现还表明，先前在真菌中未发现的一类驱动蛋白马达实际上可能在这类生物体中发挥作用。
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