Characterization of kinesin switch I mutations that cause hereditary spastic paraplegia.
Characterization of kinesin switch I mutations that cause hereditary spastic paraplegia.
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DOI：
10.1371/journal.pone.0180353
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Huckaba TM
中科院分区：
文献类型：
--
作者：
Jennings S;Chenevert M;Liu L;Mottamal M;Wojcik EJ;Huckaba TM
Kif 5A是细胞运输马达的驱动蛋白-1家族的神经元富集的同种型。在患有复杂形式的遗传性痉挛性截瘫（HSP）的患者中，已经确定了Kif 5A运动结构域中的23个单独的突变。我们对在开关I结构域中具有引起HSP的突变的二聚体重组Kif 5A进行了体外测定，开关I结构域参与驱动蛋白对ATP的协调和水解。我们观察到各种显着降低的催化和机械活动的结果，每个突变，与共享的表型，从每个运动性显着降低。在我们的反应缓冲液中用Mn 2+取代Mg 2+，在S203 C突变体的催化和整体机械性能方面提供了剂量依赖性的拯救。这项工作为具有这些突变的患者中HSP的原因提供了机理上的见解，并指出了未来的实验，以进一步剖析这种疾病的根本原因。
Kif5A is a neuronally-enriched isoform of the Kinesin-1 family of cellular transport motors. 23 separate mutations in the motor domain of Kif5A have been identified in patients with the complicated form of hereditary spastic paraplegia (HSP). We performed in vitro assays on dimeric recombinant Kif5A with HSP-causing mutations in the Switch I domain, which participates in the coordination and hydrolysis of ATP by kinesin. We observed a variety of significantly reduced catalytic and mechanical activities as a result of each mutation, with the shared phenotype from each that motility was significantly reduced. Substitution of Mn2+ for Mg2+ in our reaction buffers provides a dose-dependent rescue in both the catalytic and ensemble mechanical properties of the S203C mutant. This work provides mechanistic insight into the cause of HSP in patients with these mutations and points to future experiments to further dissect the root cause of this disease.