Structural and functional insight into ADF/cofilin from Trypanosoma brucei.

Structural and functional insight into ADF/cofilin from Trypanosoma brucei.
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从布氏锥虫中了解 ADF/Cofilin 的结构和功能

DOI:
10.1371/journal.pone.0053639
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发表时间:
2013
期刊:
影响因子:
3.7
通讯作者:
Tu X
Tu X
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Dai K;Liao S;Zhang J;Zhang X;Tu X

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ADF/cofilin家族被认为是一组肌动蛋白结合蛋白,对控制细胞内肌动蛋白的组装至关重要。本研究采用核磁共振光谱法测定了布鲁氏锥虫ADF/cofilin的溶液结构。TbCof采用保守的ADF/cofilin折叠,其中心β-片由6条β-链和5条α-螺旋组成。等温滴定量热实验表明,TbCof与g -肌动蛋白具有亚微摩尔亲和力,在低离子强度下,TbCof与adp - g -肌动蛋白的亲和力比TbCof与atp - g -肌动蛋白的亲和力高5倍。电镜和肌动蛋白丝沉降实验结果表明,TbCof解聚但不与肌动蛋白丝共沉降,其f -肌动蛋白解聚能力与pH无关。与肌动蛋白相似,TbCof分布在细胞质中。我们所有的数据表明,在结构和功能上保守的ADF/cofilin来自布鲁氏锥虫。
The ADF/cofilin family has been characterized as a group of actin-binding proteins critical for controlling the assembly of actin within the cells. In this study, the solution structure of the ADF/cofilin from Trypanosoma brucei (TbCof) was determined by NMR spectroscopy. TbCof adopts the conserved ADF/cofilin fold with a central β-sheet composed of six β-strands surrounded by five α-helices. Isothermal titration calorimetry experiments denoted a submicromolar affinity between TbCof and G-actin, and the affinity between TbCof and ADP-G-actin was five times higher than that between TbCof and ATP-G-actin at low ionic strength. The results obtained from electron microscopy and actin filament sedimentation assays showed that TbCof depolymerized but did not co-sediment with actin filaments and its ability of F-actin depolymerization was pH independent. Similar to actin, TbCof was distributed throughout the cytoplasm. All our data indicate a structurally and functionally conserved ADF/cofilin from Trypanosoma brucei.
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