Mitchell David R, Smith Brandon
Department of Cell and Developmental Biology, SUNY Upstate Medical University, Syracuse, New York, USA.
Methods Cell Biol. 2009;92:197-213. doi: 10.1016/S0091-679X(08)92013-6. Epub 2009 Nov 21.
The central pair microtubule complex in Chlamydomonas flagella has been well characterized as a regulator of flagellar dynein activity, but many aspects of this regulation depend on specific interactions between the asymmetric central pair structure and radial spokes, which appear symmetrically arranged along all nine outer doublet microtubules. Relationships between central pair-radial spoke interactions and dynein regulation have been difficult to understand because the Chlamydomonas central pair is twisted in vivo and rotates during bend propagation. Here we describe genetic and biochemical methods of dissecting the Chlamydomonas central pair and electron microscopic methods useful to determine structure-function relationships in this complex.
衣藻鞭毛中的中央微管对复合体已被充分表征为鞭毛动力蛋白活性的调节剂,但这种调节的许多方面取决于不对称中央对结构与径向辐条之间的特定相互作用,而径向辐条似乎沿所有九条外双联微管对称排列。中央对-径向辐条相互作用与动力蛋白调节之间的关系一直难以理解,因为衣藻中央对在体内是扭曲的,并在弯曲传播过程中旋转。在这里,我们描述了剖析衣藻中央对的遗传和生化方法,以及有助于确定该复合体结构-功能关系的电子显微镜方法。