Transplantation of a 17-amino acid alpha-helical DNA-binding domain into an antibody molecule confers sequence-dependent DNA recognition.
Transplantation of a 17-amino acid alpha-helical DNA-binding domain into an antibody molecule confers sequence-dependent DNA recognition.
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将 17 个氨基酸的 α 螺旋 DNA 结合结构域移植到抗体分子中可实现序列依赖性 DNA 识别。
DOI：
10.1073/pnas.92.11.5214
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发表时间：
1995
影响因子：
11.1
通讯作者：
P. Ghazal
中科院分区：
文献类型：
--
作者：
K. E. Mclane;D. Burton;P. Ghazal
能够与核酸序列特异性相互作用的重组抗体是一类DNA和rna结合蛋白，在基础研究和医学领域具有广泛的应用潜力。我们描述了一种dna结合抗体Fab-Ebox的合理设计，通过用来自B类碱性螺旋-环-螺旋蛋白TFEB的17个氨基酸结构域取代免疫球蛋白重链的可变片段。通过电泳迁移位移试验研究了DNA结合活性，结果显示Fab-Ebox与含有TFEB识别基序（CACGTG）的DNA形成特异性复合物。TFEB和Fab-Ebox在区分识别序列改变的寡脱氧核糖核苷酸方面的能力相似。胞嘧啶残基甲基化对结合的干扰表明，Fab-Ebox和TFEB都是通过沿双链DNA主槽的相互作用与DNA接触的。本研究结果表明，利用免疫球蛋白和转录因子的模块化特性，可以开发出高特异性的dna结合抗体。
Recombinant antibodies capable of sequence-specific interactions with nucleic acids represent a class of DNA- and RNA-binding proteins with potential for broad application in basic research and medicine. We describe the rational design of a DNA-binding antibody, Fab-Ebox, by replacing a variable segment of the immunoglobulin heavy chain with a 17-amino acid domain derived from TFEB, a class B basic helix-loop-helix protein. DNA-binding activity was studied by electrophoretic mobility-shift assays in which Fab-Ebox was shown to form a specific complex with DNA containing the TFEB recognition motif (CACGTG). Similarities were found in the abilities of TFEB and Fab-Ebox to discriminate between oligodeoxyribonucleotides containing altered recognition sequences. Comparable interference of binding by methylation of cytosine residues indicated that Fab-Ebox and TFEB both contact DNA through interactions along the major groove of double-stranded DNA. The results of this study indicate that DNA-binding antibodies of high specificity can be developed by using the modular nature of both immunoglobulins and transcription factors.
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影响因子：
56.9
作者：
EPHRUSSI, A;CHURCH, GM;GILBERT, W
通讯作者：
GILBERT, W
影响因子：
56.9
作者：
BLACKWELL, TK;KRETZNER, L;WEINTRAUB, H
通讯作者：
WEINTRAUB, H
二氢胸腺嘧啶（DNA 的放射分解产物）抗体的表征。
影响因子：
2.9
作者：
Hubbard,K;Ide,H;Erlanger,BF;Wallace,SS
通讯作者：
Wallace,SS
影响因子：
3.5
作者：
PARMLEY, SF;SMITH, GP
通讯作者：
SMITH, GP
影响因子：
3.5
作者：
HORTON, RM;HUNT, HD;PEASE, LR
通讯作者：
PEASE, LR