Isolation and mapping of self-assembling protein domains encoded by the Saccharomyces cerevisiae genome using lambda repressor fusions.
Isolation and mapping of self-assembling protein domains encoded by the Saccharomyces cerevisiae genome using lambda repressor fusions.
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使用 lambda 阻遏物融合对酿酒酵母基因组编码的自组装蛋白结构域进行分离和定位。
DOI：
10.1002/yea.867
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Hu,JamesC
中科院分区：
文献类型：
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作者：
Mariño-Ramírez,Leonardo;Hu,JamesC
从蛋白质结构和功能的角度来看，了解蛋白质如何形成稳定的复合物具有根本意义。在这里，我们表明，通过选择能够驱动噬菌体 λ 阻遏物 DNA 结合结构域组装的多肽，λ 阻遏物融合可用于鉴定和表征由酿酒酵母基因组编码的同型相互作用结构域。通过克隆S的随机片段构建了三个高复杂性文库。 cerevisiaeDNA 作为 λ 阻遏物融合。编码同型相互作用的阻遏蛋白融合体被恢复，识别出 35 种酵母蛋白中的寡聚单元。其中 17 个相互作用域以前未曾报道过，而另外 18 个代表同型相互作用，已在不同细节水平上进行了表征。新颖的相互作用包括几个预测的卷曲螺旋以及未知结构的域。随着基因组序列的可用性，应该可以应用这种方法，该方法提供了有关蛋白质-蛋白质相互作用的信息，与酵母双杂交筛选获得的信息互补，在酵母或其他无法获得大规模蛋白质-蛋白质相互作用数据的生物体的基因组范围内。版权所有 © 2002 约翰威利父子有限公司
Understanding how proteins are able to form stable complexes is of fundamental interest from the perspective of protein structure and function. Here we show that λ repressor fusions can be used to identify and characterize homotypic interaction domains encoded by the genome ofSaccharomyces cerevisiae, using a selection for polypeptides that can drive the assembly of the DNA binding domain of bacteriophage λ repressor. Three high complexity libraries were constructed by cloning random fragments ofS. cerevisiaeDNA as λ repressor fusions. Repressor fusions encoding homotypic interactions were recovered, identifying oligomerization units in 35 yeast proteins. Seventeen of these interaction domains have not been previously reported, while the other 18 represent homotypic interactions that have been characterized at varying levels of detail. The novel interactions include several predicted coiled‐coils as well as domains of unknown structure. With the availability of genomic sequences it should be possible to apply this approach, which provides information about protein–protein interactions that is complementary to that obtained from yeast two‐hybrid screens, on a genome‐wide scale in yeast or other organisms where large‐scale protein–protein interaction data is not available. Copyright © 2002 John Wiley & Sons, Ltd.