METHODS FOR STRUCTURAL DETERMINATION OF SELENIUM DERIVATIZED NUCLEIC ACID COMPLEXES
First Claim
1. A method of crystallizing a molecule of interest in complex with nucleic acid, comprising the steps of(a) contacting the molecule of interest with selenium-derivatized nucleic acid;
- and(b) crystallizing the molecule of interest/selenium-derivatized nucleic acid complex.
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Abstract
Methods for crystallizing a molecule of interest, such as a polypeptide, in complex with nucleic acid, including contacting the molecule of interest with selenium-derivatized nucleic acid and crystallizing the molecule of interest/selenium-derivatized nucleic acid complex are provided. Methods for determining the X-ray crystal structure of molecule of interest/selenium-derivatized nucleic acid complexes are also provided. Typically, the method of X-ray crystal structural determination includes selenium single-wavelength anomalous phasing of the selenium-derivatized nucleic acid. In some embodiments the phases for the X-ray crystal structure of the molecule of interest are not provided from another crystal. Also disclosed are methods of affecting a biological process by administering a functional nucleic acid to a cell or a subject and/or by bringing into contact a nuclease and a functional nucleic acid, where the functional nucleic acid is selenium-derivatized nucleic acid.
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Citations
29 Claims
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1. A method of crystallizing a molecule of interest in complex with nucleic acid, comprising the steps of
(a) contacting the molecule of interest with selenium-derivatized nucleic acid; - and
(b) crystallizing the molecule of interest/selenium-derivatized nucleic acid complex. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 17, 18, 19)
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9. A method of determining the interaction between a molecule of interest and nucleic acid, comprising the steps of
(a) contacting the molecule of interest with a selenium-derivatized nucleic acid; -
(b) crystallizing the molecule of interest/selenium-derivatized nucleic acid complex; (c) determining the three dimensional X-ray crystal structure of the molecule of interest/selenium-derivatized nucleic acid complex, wherein the method of X-ray crystal structural determination comprises selenium single-wavelength anomalous phasing of the selenium-derivatized nucleic acid; and (d) identifying the interfacing elements between the molecule of interest and selenium-derivatized nucleic acid in the three dimensional X-ray crystal structure of the molecule of interest/selenium-derivatized nucleic acid complex. - View Dependent Claims (10, 11, 12, 16)
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13. A method of selecting crystals containing selenium-derivatized nucleic acid, comprising the steps of
(a) growing crystals in a solution containing selenium-derivatized nucleic acid; (b) selecting crystals that are colored yellow, indicating the presence of selenium-derivatized nucleic acid. - View Dependent Claims (14, 15)
- 20. A complex of a molecule of interest and a selenium-derivatized nucleic acid.
- 22. A method of affecting a biological process, the method comprising administering a functional nucleic acid to a cell or a subject, wherein the functional nucleic acid is selenium-derivatized nucleic acid.
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25. A method of affecting a biological process, the method comprising bringing into contact a nuclease and a functional nucleic acid, wherein the functional nucleic acid is selenium-derivatized nucleic acid.
Specification