Department of Biological Sciences and the Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180.
Department of Biological Sciences and the Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York 12180
J Biol Chem. 2019 Dec 27;294(52):20070-20083. doi: 10.1074/jbc.RA119.010725. Epub 2019 Nov 20.
Heterodimeric kinesin family member KIF3AC is a mammalian kinesin-2 that is highly expressed in the central nervous system and has been implicated in intracellular transport. KIF3AC is unusual in that the motility characteristics of KIF3C when expressed as a homodimer are exceeding slow, whereas homodimeric KIF3AA, as well as KIF3AC, have much faster ATPase kinetics and single molecule velocities. Heterodimeric KIF3AC and homodimeric KIF3AA and KIF3CC are processive, although the run length of KIF3AC exceeds that of KIF3AA and KIF3CC. KIF3C is of particular interest because it exhibits a signature 25-residue insert of glycine and serine residues in loop L11 of its motor domain, and this insert is not present in any other kinesin, suggesting that it confers specific properties to mammalian heterodimeric KIF3AC. To gain a better understanding of the mechanochemical potential of KIF3AC, we pursued a single molecule study to characterize the navigation ability of KIF3AC, KIF3AA, and KIF3CC when encountering microtubule intersections. The results show that all three motors exhibited a preference to remain on the same microtubule when approaching an intersection from the top microtubule, and the majority of track switches occurred from the bottom microtubule onto the top microtubule. Heterodimeric KIF3AC and homodimeric KIF3AA displayed a similar likelihood of switching tracks (36.1 and 32.3%, respectively). In contrast, KIF3CC detached at intersections (67.7%) rather than switch tracks. These results indicate that it is the properties of KIF3A that contribute largely to the ability of KIF3AC to switch microtubule tracks to navigate intersections.
异源二聚体驱动蛋白家族成员 KIF3AC 是一种在中枢神经系统中高度表达的哺乳动物驱动蛋白-2,它与细胞内运输有关。KIF3AC 的运动特性非常特殊,当它作为同源二聚体表达时,其运动速度非常缓慢,而同源二聚体 KIF3AA 以及 KIF3AC 的 ATP 酶动力学和单分子速度则要快得多。异源二聚体 KIF3AC 和同源二聚体 KIF3AA 和 KIF3CC 都是连续的,尽管 KIF3AC 的运行长度超过了 KIF3AA 和 KIF3CC。KIF3C 特别有趣,因为它在其马达结构域的 L11 环中具有 25 个甘氨酸和丝氨酸残基的特征插入片段,而在其他任何驱动蛋白中都不存在,这表明它赋予了哺乳动物异源二聚体 KIF3AC 特定的性质。为了更好地了解 KIF3AC 的机械化学潜力,我们进行了一项单分子研究,以表征 KIF3AC、KIF3AA 和 KIF3CC 在遇到微管交叉时的导航能力。结果表明,当从顶部微管接近交叉点时,所有三种马达都表现出优先保持在同一微管上的趋势,并且大多数轨道切换发生在底部微管到顶部微管上。异源二聚体 KIF3AC 和同源二聚体 KIF3AA 显示出相似的轨道切换可能性(分别为 36.1%和 32.3%)。相比之下,KIF3CC 在交叉点处脱离(67.7%)而不是切换轨道。这些结果表明,是 KIF3A 的特性在很大程度上决定了 KIF3AC 切换微管轨道以导航交叉点的能力。