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PDBsum entry 1vro
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References listed in PDB file
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Key reference
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Title
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Selenium-Assisted nucleic acid crystallography: use of phosphoroselenoates for mad phasing of a DNA structure.
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Authors
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C.J.Wilds,
R.Pattanayek,
C.Pan,
Z.Wawrzak,
M.Egli.
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Ref.
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J Am Chem Soc, 2002,
124,
14910-14916.
[DOI no: ]
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PubMed id
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Abstract
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The combination of synchrotron radiation and a variety of atoms or ions (either
covalently attached to the biomolecule prior to crystallization or soaked into
crystals) that serve as anomalous scatterers constitutes a powerful tool in the
X-ray crystallographer's repertoire of structure determination techniques.
Phosphoroselenoates in which one of the nonbridging phosphate oxygens in the
backbone is replaced by selenium offer a simplified means for introducing an
anomalous scatterer into oligonucleotides by conventional solid-phase synthesis.
Unlike other methods that are used to derivatize DNA or RNA by covalent
attachment of a heavy atom (i.e., bromine at the C5 position of pyrimidines),
tedious synthesis of specialized nucleosides is not required. Introduction of
selenium is readily accomplished in solid-phase oligonucleotide synthesis by
replacing the standard oxidation agent with a solution of potassium
selenocyanide. This results in a diastereomeric mixture of phosphoroselenoates
that can be separated by strong anion-exchange HPLC. As a test case, all 10 DNA
hexamers of the sequence CGCGCG containing a single phosphoroselenoate linkage
(PSe) were prepared. Crystals were grown for a subset of them, and the structure
of [d(C(PSe)GCGCG)](2) was determined by the multiwavelength anomalous
dispersion technique and refined to 1.1 A resolution.
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Headers
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