利用联吡啶氨基酸对铁（II）-指状肽进行进化。
Evolution of iron(II)-finger peptides by using a bipyridyl amino acid.
作者信息
Kang Mingchao, Light Kenneth, Ai Hui-Wang, Shen Weijun, Kim Chan Hyuk, Chen Peng R, Lee Hyun Soo, Solomon Edward I, Schultz Peter G
机构信息
Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Rd, La Jolla, CA 92037.
Department of Chemistry, Stanford University, 333 Campus Drive, Stanford CA 94305.
出版信息
Chembiochem. 2014 Apr 14;15(6):822-825. doi: 10.1002/cbic.201300727. Epub 2014 Mar 3.
Abstract
We report the engineering of zinc-finger-like motifs containing the unnatural amino acid (2,2'-bipyridin-5-yl)alanine (Bpy-Ala). A phage-display library was constructed in which five residues in the N-terminal finger of zif268 were randomized to include both canonical amino acids and Bpy-Ala. Panning of this library against a nine-base-pair DNA binding site identified several Bpy-Ala-containing functional Zif268 mutants. These mutants bind the Zif268 recognition site with affinities comparable to that of the wild-type protein. Further characterization indicated that the mutant fingers bind low-spin Fe(II) rather than Zn(II) . This work demonstrates that an expanded genetic code can lead to new metal ion binding motifs that can serve as structural, catalytic, or regulatory elements in proteins.
摘要
我们报道了含有非天然氨基酸（2,2'-联吡啶-5-基）丙氨酸（Bpy-Ala）的锌指样基序的工程化。构建了一个噬菌体展示文库，其中zif268 N端指中的五个残基被随机化，以包括天然氨基酸和Bpy-Ala。针对一个九碱基对的DNA结合位点对该文库进行淘选，鉴定出了几个含有Bpy-Ala的功能性Zif268突变体。这些突变体以与野生型蛋白相当的亲和力结合Zif268识别位点。进一步的表征表明，突变指结合低自旋Fe(II)而非Zn(II)。这项工作表明，扩展的遗传密码可以导致新的金属离子结合基序，这些基序可作为蛋白质中的结构、催化或调节元件。
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