SYNZIP protein interaction toolbox: in vitro and in vivo specifications of heterospecific coiled-coil interaction domains.
SYNZIP protein interaction toolbox: in vitro and in vivo specifications of heterospecific coiled-coil interaction domains.
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DOI：
10.1021/sb200015u
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发表时间：
2012-04-20
影响因子：
4.7
通讯作者：
Keating, Amy E.
中科院分区：
文献类型：
--
作者：
Thompson, Kenneth Evan;Bashor, Caleb J.;Lim, Wendell A.;Keating, Amy E.
合成生物学工具包包含越来越多的调节转录和翻译的部分，但很少有可用于控制蛋白质缔合的部分。在这里，我们报告表征22个以前发表的异源特异性合成卷曲螺旋肽称为SYNZIPs。我们目前的生物物理分析的寡聚化状态，螺旋方向，和亲和力的27 SYNZIP对。还在两个基于细胞的测定中测试了SYNZIP对的相互作用。在酵母双杂交筛选中，253种可比较的相互作用中>85%与先前使用卷曲螺旋微阵列进行的体外测量一致。在由卷曲螺旋介导的支架控制的酵母信号传导测定中，12个SYNZIP对被成功地用于在用α因子处理后下调报告基因的表达。这些相互作用模块的表征显着增加了合成生物学可用的蛋白质相互作用部分的数量，并应促进广泛的分子工程应用。27个SYNZIP肽对的总结特征见支持信息和SYNZIP网站[]中的质量标准表。
The synthetic biology toolkit contains a growing number of parts for regulating transcription and translation, but very few that can be used to control protein association. Here we report characterization of 22 previously published heterospecific synthetic coiled-coil peptides called SYNZIPs. We present biophysical analysis of the oligomerization states, helix orientations, and affinities of 27 SYNZIP pairs. SYNZIP pairs were also tested for interaction in two cell-based assays. In a yeast two-hybrid screen, >85% of 253 comparable interactions were consistent with prior in vitro measurements made using coiled-coil microarrays. In a yeast-signaling assay controlled by coiled-coil mediated scaffolding, 12 SYNZIP pairs were successfully used to down-regulate the expression of a reporter gene following treatment with α-factor. Characterization of these interaction modules dramatically increases the number of available protein interaction parts for synthetic biology and should facilitate a wide range of molecular engineering applications. Summary characteristics of 27 SYNZIP peptide pairs are reported in specification sheets available in the Supporting Information and at the SYNZIP Web site [].
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影响因子：
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作者：
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