Párraga G, Horvath S J, Eisen A, Taylor W E, Hood L, Young E T, Klevit R E
Department of Biochemistry, University of Washington, Seattle 98195.
Science. 1988 Sep 16;241(4872):1489-92. doi: 10.1126/science.3047872.
In the proposed "zinc finger" DNA-binding motif, each repeat unit binds a zinc metal ion through invariant Cys and His residues and this drives the folding of each 30-residue unit into an independent nucleic acid-binding domain. To obtain structural information, we synthesized single and double zinc finger peptides from the yeast transcription activator ADR1, and assessed the metal-binding and DNA-binding properties of these peptides, as well as the solution structure of the metal-stabilized domains, with the use of a variety of spectroscopic techniques. A single zinc finger can exist as an independent structure sufficient for zinc-dependent DNA binding. An experimentally determined model of the single finger is proposed that is consistent with circular dichroism, one- and two-dimensional nuclear magnetic resonance, and visual spectroscopy of the single-finger peptide reconstituted in the presence of zinc.
在提出的“锌指”DNA结合基序中,每个重复单元通过不变的半胱氨酸(Cys)和组氨酸(His)残基结合一个锌金属离子,这促使每个30个残基的单元折叠成一个独立的核酸结合结构域。为了获得结构信息,我们从酵母转录激活因子ADR1合成了单锌指和双锌指肽,并使用多种光谱技术评估了这些肽的金属结合和DNA结合特性,以及金属稳定结构域的溶液结构。单个锌指可以作为一种独立结构存在,足以实现依赖锌的DNA结合。提出了一个通过实验确定的单指模型,该模型与圆二色性、一维和二维核磁共振以及在锌存在下重构的单指肽的可见光谱一致。