Methods in Enzymology Vol.59 Nucleic Acids and Protein Synthesis, Part G

This chapter describes the procedures involved in the determination of a crystal structure of a tRNA by crystallographic method. Determination of the crystal structure of a transfer RNA involves four principal steps: (a) crystallization of the molecule, (b) collection of X-ray diffraction data from native and heavy-atom derivative crystals, (c) construction and interpretation of electron-density maps, and (d) refinement of the crystal structure. Of these four steps, the art of crystallization is the least understood, but probably the most important; indeed, it is often termed as the “rate-limiting” step in structural determination. The chapter describes how various transfer RNAs are crystallized because there is no one general method for obtaining single crystals suitable for X-ray diffraction. Technical details, density measurement of crystals, and crystal mounting are also described. An optical analogy is used to describe X-ray diffraction phenomena, the multiple isomorphous replacement method, and construction and refinement of molecular models from the diffraction data. The structural features of transfer RNA that are considered reliably determined and functionally significant are summarized.

• Hardcover: 939 pages
• Publisher: Academic Press;
• Language: English
• ISBN-10: 0121819590
• ISBN-13: 978-0121819590
• Product Dimensions: 6 x 9 inches

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