Head and Rod 1 Interactions in Vimentin

Head and Rod 1 Interactions in Vimentin
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波形蛋白中头和杆 1 的相互作用

DOI:
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发表时间:
2009
影响因子:
4.8
通讯作者:
J. Voss
J. Voss
中科院分区:
生物学2区
文献类型:
--
作者:
A. Aziz;J. Hess;Madhu S. Budamagunta;P. Fitzgerald;J. Voss

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我们使用定点自旋标记 (SDSL) 和电子顺磁共振 (EPR) 来鉴定残基 17 和 137 作为中间丝蛋白波形蛋白的头结构域和杆结构域 1A 之间的相互作用位点。与位于头和杆区域的上游和下游位置的自旋标记相比,这种相互作用是最大的,表明残基 17 和 137 是该区域中最近的相互作用点。残基 120-145 的 SDSL EPR 表征(包括与杆 1A 的头部接触位点)揭示该区域表现出指示 α-螺旋卷曲螺旋结构的七肽重复模式,但该七肽重复模式在残基 139 附近开始衰减,表明卷曲螺旋结构的转变。通过监测体外组装过程中位于 17 和 137 残基处的自旋标签的光谱,我们发现 17-137 相互作用发生在组装过程的早期。我们还探讨了磷酸化对17-137相互作用的影响,发现磷酸化诱导的变化影响二聚体中的头-头相互作用(17-17),但没有显着影响二聚体中的杆-杆(137-137)和头-杆(17-137)相互作用。这些数据提供了组装中间丝中头杆相互作用的第一个直接证据和位置,以及杆 1A 中卷曲线圈结构的直接证据。最后,数据确定了体外磷酸化后该区域结构的变化。
We have used site-directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) to identify residues 17 and 137 as sites of interaction between the head domain and rod domain 1A of the intermediate filament protein vimentin. This interaction was maximal when compared with the spin labels placed at up- and downstream positions in both head and rod regions, indicating that residues 17 and 137 were the closest point of interaction in this region. SDSL EPR characterization of residues 120-145, which includes the site of head contact with rod 1A, reveals that this region exhibits the heptad repeat pattern indicative of α-helical coiled-coil structure, but that this heptad repeat pattern begins to decay near residue 139, suggesting a transition out of coiled-coil structure. By monitoring the spectra of spin labels placed at the 17 and 137 residues during in vitro assembly, we show that 17-137 interaction occurs early in the assembly process. We also explored the effect of phosphorylation on the 17-137 interaction and found that phosphorylation-induced changes affected the head-head interaction (17-17) in the dimer, without significantly influencing the rod-rod (137-137) and head-rod (17-137) interactions in the dimer. These data provide the first direct evidence for, and location of, headrod interactions in assembled intermediate filaments, as well as direct evidence of coiled-coil structure in rod 1A. Finally, the data identify changes in the structure in this region following in vitro phosphorylation.
通过定点自旋标记和电子顺磁共振鉴定波形蛋白中间丝中磷酸化诱导的变化。
DOI: 10.1021/bi801137m
发表时间: 2008
期刊: Biochemistry
影响因子: 2.9
作者:
Pittenger,JoshT;Hess,JohnF;Budamagunta,MadhuS;Voss,JohnC;Fitzgerald,PaulG
通讯作者: Fitzgerald,PaulG
DOI: 10.1073/pnas.92.18.8259
发表时间: 1995-08-29
影响因子: 11.1
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DOI: 10.1016/j.jmb.2004.04.039
发表时间: 2004-06-25
影响因子: 5.6
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通讯作者: Herrmann, H
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DOI: 10.1046/j.1432-1327.2001.01946.x
发表时间: 2001
期刊: European journal of biochemistry
影响因子: --
作者:
Chomiki,N;Voss,JC;Warden,CH
通讯作者: Warden,CH
DOI: 10.1016/j.yexcr.2007.04.007
发表时间: 2007
影响因子: 3.7
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通讯作者: Oshima,RobertG