Selective recognition of a DNA G-quadruplex by an engineered antibody.
Selective recognition of a DNA G-quadruplex by an engineered antibody.
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DOI：
10.1021/bi800983u
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发表时间：
2008-09-09
期刊：
影响因子：
2.9
通讯作者：
Balasubramanian, Shankar
中科院分区：
文献类型：
--
作者：
Fernando, Himesh;Rodriguez, Raphael;Balasubramanian, Shankar
特定的富含鸟嘌呤的核酸序列可以折叠成称为G-四链体的稳定二级结构。这些结构已在基因组的各个区域中被识别，包括端粒、基因启动子和 UTR 区域，这增加了它们可能与生物功能相关的可能性。计算分析预测，分子内 G-四链体形成序列在人类基因组中普遍存在，从而提高了差异识别基因组 G-四链体的愿望。我们采用抗体噬菌体展示和竞争选择技术来生成单链抗体，该抗体在 G 四链体和双链体 DNA 之间显示出 > 1000 倍的辨别力，此外，在两个相关的分子内平行 DNA G 四链体之间显示出 > 100 倍的辨别力。抗原结合位点的氨基酸序列组成显示所选 scFv 的轻链和重链内的保守性，表明 G-四链体识别的序列要求。圆二色性(CD)光谱数据表明scFv与预折叠的G-四链体结合并且不诱导G-四链体结构形成。这项研究证明了我们所知道的两个分子内基因组 G 四链体之间最强的区别。
Particular guanine rich nucleic acid sequences can fold into stable secondary structures called G-quadruplexes. These structures have been identified in various regions of the genome that include the telomeres, gene promoters and UTR regions, raising the possibility that they may be associated with biological function(s). Computational analysis has predicted that intramolecular G-quadruplex forming sequences are prevalent in the human genome, thus raising the desire to differentially recognize genomic G-quadruplexes. We have employed antibody phage display and competitive selection techniques to generate a single-chain antibody that shows >1000-fold discrimination between G-quadruplex and duplex DNA, and furthermore >100-fold discrimination between two related intramolecular parallel DNA G-quadruplexes. The amino acid sequence composition at the antigen binding site shows conservation within the light and heavy chains of the selected scFvs, suggesting sequence requirements for G-quadruplex recognition. Circular dichroism (CD) spectroscopic data showed that the scFv binds to the prefolded G-quadruplex and does not induce G-quadruplex structure formation. This study demonstrates the strongest discrimination that we are aware of between two intramolecular genomic G-quadruplexes.
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影响因子：
2.9
作者：
Ladame, S;Schouten, JA;Roldan, J;Redman, JE;Neidle, S;Balasubramanian, S
通讯作者：
Balasubramanian, S
TrisubStttestated的异丙恶唑作为新的G四链体结合配体：C-KIT癌基因表达的小分子调节。
影响因子：
15
作者：
Bejugam, Mallesham;Sewitz, Sven;Shirude, Pravin S.;Rodriguez, Raphael;Shahid, Ramla;Balasubramanian, Shankar
通讯作者：
Balasubramanian, Shankar
影响因子：
46.9
作者：
de Wildt, RMT;Mundy, CR;Tomlinson, IM
通讯作者：
Tomlinson, IM
影响因子：
2.9
作者：
Isalan, M;Patel, SD;Choo, Y
通讯作者：
Choo, Y
DOI：
10.1073/pnas.89.18.8832
发表时间：
1992-09-15
影响因子：
11.1
作者：
JIN, RZ;GAFFNEY, BL;BRESLAUER, KJ
通讯作者：
BRESLAUER, KJ