Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport.
Preferential binding of a kinesin-1 motor to GTP-tubulin-rich microtubules underlies polarized vesicle transport.
复制标题
DOI：
10.1083/jcb.201104034
复制
发表时间：
2011-07-25
期刊：
影响因子：
--
通讯作者：
Hirokawa N
中科院分区：
文献类型：
--
作者：
Nakata T;Niwa S;Okada Y;Perez F;Hirokawa N
KIF 5对富含GTP-微管蛋白的微管的高亲和力导致极化运动蛋白在神经元轴突尖端的积累，并且可能是极化囊泡运输的基础。神经元中的极化运输是神经元回路形成的基础。含有马达结构域的截短型KIF 5（一种驱动蛋白-1）识别富含EB 1结合位点的轴突微管，并选择性地聚集在轴突尖端。然而，它仍然是未知的提示KIF 5识别导致这种选择性积累。我们发现，轴突微管优先染色的抗GTP-微管蛋白抗体hMB 11。超分辨率显微镜结合EM免疫细胞化学显示hMB 11定位于KIF 5的附着位点。此外，EB 1，它优先结合鸟苷酰亚甲基二磷酸（GMPCPP）微管在体外，识别轴突微管上的hMB 11结合位点。此外，hMB 11抗体在神经元中的表达破坏了截短的KIF 5在轴突尖端的选择性积累。体外研究显示，KIF 5马达头与GMPCPP微管的结合比与GDP微管的结合强约三倍。总的来说，这些数据表明，丰富的GTP-微管蛋白的轴突微管可能是选择性KIF 5本地化和极化轴突囊泡运输的基础。
The high affinity of KIF5 for microtubules rich in GTP-tubulin results in polarized motor protein accumulation at axonal tips in neurons and may underlie polarized vesicle transport. Polarized transport in neurons is fundamental for the formation of neuronal circuitry. A motor domain–containing truncated KIF5 (a kinesin-1) recognizes axonal microtubules, which are enriched in EB1 binding sites, and selectively accumulates at the tips of axons. However, it remains unknown what cue KIF5 recognizes to result in this selective accumulation. We found that axonal microtubules were preferentially stained by the anti–GTP-tubulin antibody hMB11. Super-resolution microscopy combined with EM immunocytochemistry revealed that hMB11 was localized at KIF5 attachment sites. In addition, EB1, which binds preferentially to guanylyl-methylene-diphosphate (GMPCPP) microtubules in vitro, recognized hMB11 binding sites on axonal microtubules. Further, expression of hMB11 antibody in neurons disrupted the selective accumulation of truncated KIF5 in the axon tips. In vitro studies revealed approximately threefold stronger binding of KIF5 motor head to GMPCPP microtubules than to GDP microtubules. Collectively, these data suggest that the abundance of GTP-tubulin in axonal microtubules may underlie selective KIF5 localization and polarized axonal vesicular transport.
登录
查看更多内容
DOI：
10.1083/jcb.200911024
发表时间：
2010-08-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Friedman JR;Webster BM;Mastronarde DN;Verhey KJ;Voeltz GK
通讯作者：
Voeltz GK
影响因子：
16.2
作者：
Jacobson, C;Schnapp, B;Banker, GA
通讯作者：
Banker, GA
DOI：
10.1073/pnas.1014758108
发表时间：
2011-03-08
影响因子：
11.1
作者：
Maurer, Sebastian P.;Bieling, Peter;Surrey, Thomas
通讯作者：
Surrey, Thomas
影响因子：
21.3
作者：
Coy, DL;Hancock, WO;Howard, J
通讯作者：
Howard, J
DOI：
10.1083/jcb.109.6.3085
发表时间：
1989-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Baas PW;Black MM;Banker GA
通讯作者：
Banker GA