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Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs

Paper ID Volume ID Publish Year Pages File Format Full-Text
21279 43214 2012 8 PDF Available
Title
Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs
Abstract

In this study, we have demonstrated an accurate and rapid small RNA analytical method with both sequence determination and detailed modification analysis by electrospray ionization-ion trap/time-of-flight mass spectrometry (ESI-IT/TOFMS). To develop this ideal method, we have examined the performance of ESI-IT/TOFMS using various chemically-synthesized model sequences of modified or unmodified microRNAs (miRNAs). The deconvoluted mass of a 22-nucleotide (nt) miRNA was obtained from a multiply charged precursor ion (MS1). The ion exhibited high mass accuracy (< 7 ppm) and high mass resolution (a value of m/Δm = 10,000) and was therefore very useful in RNA composition assignment. The optimized MS2 method using ion trap collision-induced dissociation, as well as automatic annotation analysis of product ions based on the accurate mass information, enabled the precise sequencing determination of intact miRNAs. Further, the detailed structural analysis of 3′-terminal modified nucleic acid in intact methylated miRNA was carried out using the MS3 capability of the hybrid IT/TOFMS. The direct infusion method also provided a high throughput and good sensitivity because the analytical time and sample concentration needed in a series of experiments with reliable data were only 3 min and 100 nM, respectively. This study provides a novel approach for characterizing the intact chemically-synthesized small RNA without chemical and enzymatic digestions and would be widely applicable for the structural analysis of complicated modified small RNAs.

Keywords
Electrospray ionization ion trap/time-of-flight mass spectrometry (ESI-IT/TOFMS); Chemically-synthesized small RNA; MicroRNA; RNA modification; Multistep MSn analysis
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Application of electrospray ionization ion trap/time-of-flight mass spectrometry for chemically-synthesized small RNAs
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 113, Issue 3, March 2012, Pages 412–419
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us