Analysis of heterodimer formation by Xklp3A/B, a newly cloned kinesin-II from Xenopus laevis

被引:32
作者
De Marco, V
Burkhard, P
Le Bot, N
Vernos, I
Hoenger, A
机构
[1] European Mol Biol Lab, Struct & Computat Biol Programme, D-69117 Heidelberg, Germany
[2] European Mol Biol Lab, Cell Biol & Cell Biophys Programme, D-69117 Heidelberg, Germany
[3] Univ Basel, Bioctr, Abt Strukturbiol, CH-4056 Basel, Switzerland
关键词
alpha-helical coiled coil; coiled-coil trigger site; heptad repeats; kinesin-II; Xklp3A/B;
D O I
10.1093/emboj/20.13.3370
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
kinesin-II motor proteins are composed of two different kinesin-like motor proteins and one cargo binding subunit, Here we report the cloning of a new member of the kinesin-II superfamily, Xklp3A from Xenopus laevis, which forms a heterodimeric complex with Xklp3B, The heterodimer formation properties between Xklp3A and B have been tested in vitro using reticulocyte lysate expression and immnnoprecipitation, To this end we produced a series of Xklp3A and B constructs of varying length and tested their propensity for heterodimer formation. We could demonstrate that, in contrast to conventional kinesin, the critical domains for heterodimer formation in Xklp3A/B are located at the C-terminal end of the stalk. Neither the neck nor the highly charged stretches after the neck region, which are typical of kinesins-II, are required for heterodimer formation, nor do they prevent homodimer formation. Dimerization is controlled by a cooperative mechanism between the C-terminal coiled-coil segments. Classical trigger sites were not identified. The critical regions for dimerization exhibit a very high degree of sequence conservation among equivalent members of the kinesin-II family.
引用
收藏
页码:3370 / 3379
页数:10
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