Oligo-Asp Tag/Zn(II) complex probe as a new pair for labeling and fluorescence imaging of proteins
Oligo-Asp Tag/Zn(II) complex probe as a new pair for labeling and fluorescence imaging of proteins
复制标题
DOI：
10.1021/ja0618604
复制
发表时间：
2006-08-16
影响因子：
15
通讯作者：
Hamachi, Itaru
中科院分区：
文献类型：
--
作者：
Ojida, Akio;Honda, Kei;Hamachi, Itaru
为了在复杂的生物系统中完成对特定蛋白质的选择性标记，需要结合到蛋白质中的肽标签和互补的小分子探针。尽管各种肽标签/探针对已经被开发出来作为蛋白质分析的分子工具，但适用于蛋白质实时成像的对的可用性仍然有限。我们现在报道了一种新的肽标签/人工探针对，由遗传可编码的寡聚天冬氨酸序列（D4标签，（D4）(n), n = 1-3）和相应的多核Zn（II）配合物（Zn(II)-DpaTyrs）组成。Zn(II)-DpaTyr探针与D4标签的强结合亲和力是多重配位键和多价效应的结果。采用等温滴定量热法定量测定。标签和探针之间的高亲和力是选择性蛋白质标记所不可缺少的，这使得该对在完整的细胞结构中用于连接三次重复D4标签(（D4)(3)）的膜结合受体蛋白的标记和荧光成像，而不会显著影响受体信号转导。
To accomplish the selective labeling of a specific protein in complicated biological systems, a peptide tag incorporated into the protein and a complementary small molecular probe are required. Although a variety of peptide tag/probe pairs have been developed as molecular tools for protein analyses, the availability of pairs suitable for real-time imaging of proteins is still limited. We now report a new peptide tag/artificial probe pair composed of a genetically encodable oligo-aspartate sequence (D4 tag, (D4)(n), n = 1-3) and the corresponding multinuclear Zn(II) complexes (Zn(II)-DpaTyrs). The strong binding affinity of the Zn(II)-DpaTyr probes with the D4 tag was a result of the multiple coordination bonds and the multivalent effect. It was measured quantitatively by isothermal titration calorimetry. The high affinity between the tag and the probe, indispensable for the selective protein labeling, enabled the pair to be used for the labeling and fluorescence imaging of a membrane-bound receptor protein tethering a triply repeated D4 tag ((D4)(3)) in an intact cell configuration without significantly affecting the receptor signal transduction.