含 3-氰基乙烯基咔唑氨基酸的 GCN4 肽与双链 DNA 的可逆光交联。
Reversible photo-cross-linking of the GCN4 peptide containing 3-cyanovinylcarbazole amino acid to double-stranded DNA.
机构信息
School of Advanced Science and Technology, Japan Advanced Institute of Science and Technology. Asahidai 1-1, Nomi, Ishikawa 923-1292, Japan.
出版信息
Org Biomol Chem. 2019 Jun 26;17(25):6277-6283. doi: 10.1039/c9ob00372j.
Interaction analysis in vivo greatly promotes the analyses and understanding of biological functions. The interaction between DNA and peptides or proteins is very important in terms of readout and amplifying information from genomic DNA. In this study, we designed and synthesized a photo-cross-linkable amino acid, l-3-cyanovinlycarbazole amino acid (l-CNVA), to double-stranded DNA. Reversible photo-cross-linking between DNA and peptides containing CNVA, having 3-cyanovinylcarbazole moieties capable of photo-cross-linking to nucleic acids, was demonstrated. As a result, it was shown that the GCN4 peptide, containing CNVA, can be photo-cross-linked to DNA, and its adduct was photo-split into the original peptide and DNA with 312 nm-irradiation. This is the first report that reversibly manipulates photo-crosslinking between double stranded DNA and peptides. In addition, this reversible photo-cross-linking, using l-CNVA, is faster and with higher yield than that using diazirine and psoralen.
体内相互作用分析极大地促进了对生物功能的分析和理解。就从基因组 DNA 中读出和放大信息而言，DNA 与肽或蛋白质之间的相互作用非常重要。在本研究中，我们设计并合成了一种光交联氨基酸，即 l-3-氰基乙烯基咔唑氨基酸（l-CNVA），使其与双链 DNA 结合。证明了含有 CNVA 的肽与 DNA 之间的可逆光交联，其中 3-氰基乙烯基咔唑部分能够与核酸发生光交联。结果表明，含有 CNVA 的 GCN4 肽可以与 DNA 光交联，其加合物在 312nm 照射下光解为原始肽和 DNA。这是首次报道可以可逆地操纵双链 DNA 和肽之间的光交联。此外，与重氮甲烷和补骨脂素相比，使用 l-CNVA 的这种可逆光交联反应更快，产率更高。
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