通过¹³C、¹⁵N双缺失和电喷雾电离傅里叶变换离子回旋共振质谱法改进寡核糖核苷酸的质量分析。

Improved mass analysis of oligoribonucleotides by 13C, 15N double depletion and electrospray ionization FT-ICR mass spectrometry.

作者信息

Xiong Ying, Schroeder Kersten, Greenbaum Nancy L, Hendrickson Christopher L, Marshall Alan G

机构信息

Institute of Molecular Biophysics, Florida State University, Tallahassee, Florida 32306, USA.

出版信息

Anal Chem. 2004 Mar 15;76(6):1804-9. doi: 10.1021/ac030299e.

DOI:10.1021/ac030299e

PMID:15018587

Abstract

13C, 15N doubly depleted 32-ribonucleotide was synthesized enzymatically by in vitro transcription from nucleoside triphosphates isolated from E. coli grown in a minimal medium containing 12C, 14N-enriched glucose and ammonium sulfate. Following purification and desalting by reversed-phase HPLC, buffer exchange with Microcon YM-3, and ethanol precipitation, electrospray ionization Fourier transform ion cyclotron resonance mass spectra revealed greatly enhanced abundance of monoisotopic ions (by a factor of approximately 100) and a narrower isotopic distribution with higher signal-to-noise ratio. The abrupt onset and high magnitude of the monoisotopic species promise to facilitate accurate mass measurement of RNA's.

摘要

通过从在含有富含(^{12}C)、(^{14}N)的葡萄糖和硫酸铵的基本培养基中生长的大肠杆菌中分离出的三磷酸核苷进行体外转录，酶法合成了(^{13}C)、(^{15}N)双贫化的32 - 核糖核苷酸。经过反相高效液相色谱法纯化和脱盐、用Microcon YM - 3进行缓冲液交换以及乙醇沉淀后，电喷雾电离傅里叶变换离子回旋共振质谱显示单同位素离子的丰度大大提高（约为100倍），同位素分布更窄，信噪比更高。单同位素物种的突然出现和高丰度有望促进RNA的精确质量测量。