Mazumder A, Chen C B, Gaynor R, Sigman D S
Department of Biological Chemistry, School of Medicine, University of California, Los Angeles 90024-1570.
Biochem Biophys Res Commun. 1992 Sep 30;187(3):1503-9. doi: 10.1016/0006-291x(92)90472-w.
The 1,10-phenanthroline-cuprous complex (OP-Cu) with hydrogen peroxide as a coreactant nicks the single-stranded loops and bulges of RNA stem-loop structures more rapidly than the double-stranded stems. This chemical nuclease is therefore a useful footprinting reagent for these regions and can be used to monitor both intramolecular and intermolecular hybridization of single-stranded domains. The formation of A-form structures characteristic of either RNA-RNA or RNA-DNA duplexes inhibits scission because it blocks the binding site of the coordination complex in single-stranded loops and not because the oxidatively sensitive hydrogens of the ribose moiety are blocked. The C-4' and C-1' hydrogens are accessible to solvent in A-structures.
以过氧化氢作为共反应物的1,10-菲咯啉-亚铜配合物(OP-Cu)切割RNA茎环结构的单链环和凸起比双链茎更快。因此,这种化学核酸酶是用于这些区域的有用的足迹试剂,可用于监测单链结构域的分子内和分子间杂交。RNA-RNA或RNA-DNA双链体特有的A-型结构的形成会抑制切割,因为它阻断了单链环中配位复合物的结合位点,而不是因为核糖部分的氧化敏感氢被阻断。在A-型结构中,C-4'和C-1'氢可与溶剂接触。