CORRELATION BETWEEN THE ATPASE AND MICROTUBULE TRANSLOCATING ACTIVITIES OF SEA-URCHIN EGG KINESIN
CORRELATION BETWEEN THE ATPASE AND MICROTUBULE TRANSLOCATING ACTIVITIES OF SEA-URCHIN EGG KINESIN
复制标题
DOI：
10.1038/328160a0
复制
发表时间：
1987-07-09
期刊：
影响因子：
64.8
通讯作者：
SCHOLEY, JM
中科院分区：
文献类型：
--
作者：
COHN, SA;INGOLD, AL;SCHOLEY, JM
几年前在鞭毛轴突中证明了ATP水解和微管运动之间的耦合1，2，随后的研究表明，相关的微管马达动力蛋白使用ATP通过类似于肌肉中肌球蛋白的跨桥机制驱动微管滑动3，4。驱动蛋白5是一种基于微管的运动蛋白，可能参与细胞器运输和有丝分裂6，以核苷酸敏感的方式结合微管5，7，8，并需要可水解的核苷酸在玻璃表面上易位微管9，10。最近，神经元驱动蛋白被证明具有微管激活的ATP酶活性11，12，尽管ATP水解和运动性之间的偶联未被证实。本文报道了以Mg-ATP为底物时，经5′-腺苷酰亚氨基二磷酸（AMPPNP）诱导的微管结合步骤或不经AMPPNP诱导的微管结合步骤制备的海胆卵驱动蛋白也具有显著的微管激活ATP酶活性。通过加入无Mg的ATP、通过用EDTA螯合Mg 2+、通过加入Na 3 VO 4或通过加入有或没有Mg 2+的AMPPNP，这种ATP酶活性以剂量依赖性方式被抑制。加入这些相同的试剂也以剂量依赖性方式抑制海胆卵驱动蛋白的微管易位活性，支持了驱动蛋白驱动的运动性与微管激活的Mg 2 +-ATP酶活性偶联的假设。
Coupling between ATP hydrolysis and microtubule movement was demonstrated several years ago in flagellar axonemes1,2and subsequent studies suggest that the relevant microtubule motor, dynein, uses ATP to drive microtubule sliding by a cross-bridge mechanism analogous to that of myosin in muscles3,4. Kinesin5, a microtubule-based motility protein which may participate in organelle transport and mitosis6, binds microtubules in a nucleo-tide-sensitive manner5,7,8, and requires hydrolysable nucleotides to translocate microtubules over a glass surface9,10. Recently, neuronal kinesin was shown to possess microtubule-activated ATPase activity11,12although coupling between ATP hydrolysis and motility was not demonstrated. Here we report that sea urchin egg kinesin, prepared either with or without a 5′-adenylyl imido-diphosphate(AMPPNP)-induced microtubule binding step, also possesses significant microtubule-activated ATPase activity when Mg-ATP is used as a substrate. This ATPase activity is inhibited in a dose-dependent manner by addition of Mg-free ATP, by chelation of Mg2+with EDTA, by addition of Na3VO4, or by addition of AMPPNP with or without Mg2+Addition of these same reagents also inhibits the microtubule-translocating activities of sea urchin egg kinesin in a dose-dependent manner, supporting the hypothesis that kinesin-driven motility is coupled to the microtubule-activated Mg2+-ATPase activity.