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Selenium modification of nucleic acids: preparation of phosphoroselenoate derivatives for crystallographic phasing of nucleic acid structures

Nature Protocols volume 2, pages 640–646 (2007)Cite this article

Abstract

This protocol describes a simplified means of introducing an anomalously scattering atom into oligonucleotides by conventional solid-phase synthesis. Replacement of a nonbridging phosphate oxygen in the backbone with selenium is practically suitable for any nucleic acid. The resulting oligonucleotide P-diastereomers can be separated using anion exchange HPLC to yield diastereomerically pure phosphoroselenoates (PSes). The total time for the synthesis and ion-exchange HPLC separation of pure PSe is approximately 60 h.

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Figure 1
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Figure 4: Matrix-assisted laser desorption/ionization—time of flight (MALDI-TOF) mass spectra of (a) 5′-6C5G4C3G2C1G-3′ (wild type) and (b) 5′-6CPSe5G4C3G2C1G-3′ (slower eluting diastereomer).

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Acknowledgements

This work was supported by the U.S. National Institutes of Health (Grant R01 GM55237 to M.E.).

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  1. Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, 37232, Tennessee, USA

    Pradeep S Pallan & Martin Egli

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  1. Pradeep S Pallan
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Correspondence to Martin Egli.

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Pallan, P., Egli, M. Selenium modification of nucleic acids: preparation of phosphoroselenoate derivatives for crystallographic phasing of nucleic acid structures. Nat Protoc 2, 640–646 (2007). https://doi.org/10.1038/nprot.2007.74

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