Investigation of Enzymatic Behavior of Benzonase/Alkaline Phosphatase in the Digestion of Oligonucleotides and DNA by ESI-LC/MS

UNCG Author/Contributor (non-UNCG co-authors, if there are any, appear on document)
Norman H. Chiu, Assistant Professor (Creator)
The University of North Carolina at Greensboro (UNCG )
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Abstract: We have developed an ion-pairing HPLC-MS method that has sufficient separation power, selectivity, and sensitivity to investigate the enzymatic behavior of benzonase/alkaline phosphatase upon digestion of oligonucleotides and DNA. Mass spectrometry revealed that this enzyme pair can nonspecifically digest oligonucleotides and DNA into fragments ranging from 2 to 10 nucleotides, i.e., sizes suitable for routine mass spectrometric measurements. Trimers, tetramers, and pentamers are the most prominent digested products. This makes benzonase/alkaline phosphatase a promising choice for DNA and DNA adduct related studies that require a nonspecific enzyme. A computer software program developed in-house was critical in automating the processing of mass spectral data. The methodology described here provides a systematic approach for evaluating the behavior of DNA-cleaving enzymes by mass spectrometry.

Additional Information

Language: English
Date: 2007
chemistry, biochemistry, DNA, enzymatic behavior, benzonase/alkaline phosphate, oligonucleotides, mass spectronomy

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