His-Tag-Mediated Dimerization of Chemoreceptors Leads to Assembly of Functional Nanoarrays.
His-Tag-Mediated Dimerization of Chemoreceptors Leads to Assembly of Functional Nanoarrays.
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DOI：
10.1021/acs.biochem.7b00698
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发表时间：
2017-11-07
期刊：
影响因子：
2.9
通讯作者：
Thompson LK
中科院分区：
文献类型：
--
作者：
Haglin ER;Yang W;Briegel A;Thompson LK
跨膜趋化性受体在细菌中以扩展的六边形阵列被膜和胞质结合伴侣（激酶CheA和偶联蛋白CheW）稳定。阵列结构和组装模型提出受体簇成二聚体的三聚体，其与一个CheA二聚体和两个CheW单体相关联以形成信号转导所需的最小“核心单元”。在体外重建化学感受器三元复合物，是均匀的和功能性的，并表现出天然的架构仍然是一个挑战。在这里，我们报告说，组氨酸标签介导的受体二聚与二价金属是足以驱动组装的nativelike功能阵列的受体胞质片段。我们的结果表明受体二聚化启动组装，并先于具有部分激酶活性的三元复合物的形成。最大激酶活性的恢复与向更大复合物的转变相吻合，表明激酶活性取决于核心单元之外的相互作用。我们假设，实现最大的活动需要构建核心单元成六边形和/或合并成扩展的晶格六边形。总的来说，最低限度地干扰His-标签介导的二聚化导致组装化学感受器阵列与天然架构，因此作为一个强大的工具，用于研究组装和机制，这种复杂的和其他多蛋白复合物。
Transmembrane chemotaxis receptors are found in bacteria in extended hexagonal arrays stabilized by the membrane and by cytosolic binding partners, the kinase CheA and coupling protein CheW. Models of array architecture and assembly propose receptors cluster into trimers of dimers that associate with one CheA dimer and two CheW monomers to form the minimal “core unit” necessary for signal transduction. Reconstructing in vitro chemoreceptor ternary complexes that are homogeneous and functional and exhibit native architecture remains a challenge. Here we report that His-tag-mediated receptor dimerization with divalent metals is sufficient to drive assembly of nativelike functional arrays of a receptor cytoplasmic fragment. Our results indicate receptor dimerization initiates assembly and precedes formation of ternary complexes with partial kinase activity. Restoration of maximal kinase activity coincides with a shift to larger complexes, suggesting that kinase activity depends on interactions beyond the core unit. We hypothesize that achieving maximal activity requires building core units into hexagons and/or coalescing hexagons into the extended lattice. Overall, the minimally perturbing His-tag-mediated dimerization leads to assembly of chemoreceptor arrays with native architecture and thus serves as a powerful tool for studying the assembly and mechanism of this complex and other multiprotein complexes.
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影响因子：
2.9
作者：
Briegel A;Wong ML;Hodges HL;Oikonomou CM;Piasta KN;Harris MJ;Fowler DJ;Thompson LK;Falke JJ;Kiessling LL;Jensen GJ
通讯作者：
Jensen GJ
DOI：
10.1016/s0021-9673(00)93969-4
发表时间：
1987-12-18
期刊：
JOURNAL OF CHROMATOGRAPHY
影响因子：
--
作者：
HOCHULI, E;DOBELI, H;SCHACHER, A
通讯作者：
SCHACHER, A
DOI：
10.1073/pnas.1104824108
发表时间：
2011-06-07
影响因子：
11.1
作者：
Li, Mingshan;Hazelbauer, Gerald L.
通讯作者：
Hazelbauer, Gerald L.
影响因子：
3
作者：
Kremer, JR;Mastronarde, DN;McIntosh, JR
通讯作者：
McIntosh, JR
DOI：
10.1073/pnas.0802868105
发表时间：
2008-08-26
影响因子：
11.1
作者：
Besschetnova, Tatiana Y.;Montefusco, David J.;Weis, Robert M.
通讯作者：
Weis, Robert M.