Wilk H E, Kecskemethy N, Schäfer K P
Nucleic Acids Res. 1982 Dec 11;10(23):7621-33. doi: 10.1093/nar/10.23.7621.
m-Aminophenylboronate-substituted agarose binds specifically RNA chains carrying a mature 5' cap. The binding occurs most effectively at pH greater than 8 and involves diester formation between the negatively charged tetrahedric boronate group and the cisdiol of the ribose of the cap. The positive charge introduced by the m7G methylation is necessary for efficient binding although two closely spaced cis-diol groups alone (as in the cap analog NADH) are sufficient for binding. Non-capped RNA (like poly (U) and rRNA) or decapped RNA are not bound. It is shown that the matrix can be used for the isolation of capped small nuclear RNA and mRNA.
间氨基苯硼酸取代的琼脂糖能特异性结合带有成熟5'帽结构的RNA链。这种结合在pH大于8时最为有效,涉及带负电荷的四面体硼酸基团与帽结构核糖的顺式二醇之间形成二酯。m7G甲基化引入的正电荷对于有效结合是必需的,尽管仅两个紧密间隔的顺式二醇基团(如帽类似物NADH中的)就足以实现结合。未加帽的RNA(如聚(U)和rRNA)或去帽的RNA不会被结合。结果表明,该基质可用于分离加帽的小核RNA和mRNA。