Selenium modification of nucleic acids: preparation of phosphoroselenoate derivatives for crystallographic phasing of nucleic acid structures.
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.
Pallan
Pradeep S
PS
Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee 37232, USA.
Egli
Martin
M
D002851:Chromatography, High Pressure Liquid; D003461:Crystallography / Q000379:methods*; D015394:Molecular Structure; D009696:Nucleic Acids / Q000737:chemistry*; D017553:Phosphorus Compounds / Q000138:chemical synthesis* / Q000737:chemistry; D012643:Selenium / Q000737:chemistry*; D018036:Selenium Compounds / Q000138:chemical synthesis* / Q000737:chemistry
D009696:Nucleic Acids; D017553:Phosphorus Compounds; D018036:Selenium Compounds; D012643:Selenium