LENGTH-ENCODED MULTIPLEX BINDING-SITE DETERMINATION - APPLICATION TO ZINC-FINGER PROTEINS
LENGTH-ENCODED MULTIPLEX BINDING-SITE DETERMINATION - APPLICATION TO ZINC-FINGER PROTEINS
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DOI：
10.1073/pnas.91.23.11099
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发表时间：
1994-11-08
影响因子：
11.1
通讯作者：
BERG, JM
中科院分区：
文献类型：
--
作者：
DESJARLAIS, JR;BERG, JM
组合文库的筛选正在成为鉴定或精制结合蛋白、酶和其他受体的配体结构的有力方法。我们描述了一个寡核苷酸库搜索程序，其中每个成员的身份是编码的寡核苷酸的长度。该编码方案允许结合位点偏好通过变性凝胶电泳的DNA长度测定来评估。我们已经应用这种方法来确定18个Cys（2）His（2）锌指结构域作为侧翼锌指的固定背景内的中心结构域的结合位点偏好。该方法的一个优点是，除了简单地确定最佳或共有配体的序列之外，还可以估计文库所有成员的相对亲和力。确定的锌指结构域特异性将是有用的模块化锌指蛋白的设计。
The screening of combinatorial libraries is becoming a powerful method for identifying or refining the structures of ligands for binding proteins, enzymes, and other receptors. We describe an oligonucleotide library search procedure in which the identity of each member is encoded in the length of olgonucleotides. This encoding scheme allows binding-site preferences to be evaluated via DNA length determination by denaturing gel electrophoresis. We have applied this method to determine the binding site preferences for 18 Cys(2)His(2) zinc finger domains as the central domain within a fixed context of flanking zinc fingers. An advantage of the method is that the relative affinities of all members of the Library can be estimated in addition to simply determining the sequence of the optimal or consensus ligand. The zinc finger domain specificities determined will be useful for modular zinc finger protein design.