Structure of tubulin at 6.5 Å and location of the taxol-binding site

Eva Nogales, Sharon Grayer Wolf, Israr A. Khan, Richard F. Ludueña, Kenneth H. Downing

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Abstract

TUBULIN, the major component of microtubules, is a heterodimer of two chains, α and β1, both of relative molecular mass 50,000 (Mr50K) and with 40-50% identity. The isotypic variety2 and conformational flexibility of tubulin have so far made it impossible to obtain crystals for X-ray work3. Structural knowledge of tubulin has been limited to about 20 Å from X-ray diffraction of oriented microtubules4, and from electron microscopy of microtubules and zinc-induced crystalline sheets in negative stain5,6. The sheets consist of protofilaments similar to those in microtubules but associated in an antiparallel arrangement7, and their two-dimensional character is ideal for high-resolution electron microscopy8,9. Here we present a three-dimensional reconstruction of tubulin to 6.5 Å resolution, obtained by electron crystallography of zinc-induced two-dimensional crystals of the protein. The α- and β-subunits appear topologically similar, in agreement with their sequence homology10. Several features can be defined in terms of secondary structure. An apparent α-helical portion, adjacent to both interdimer and inter-protofilament contacts, is tentatively attributed to a segment near the carboxy terminus of the protein. We can assign the α- and β-subunits on the basis of projection studies of the binding of taxol*, which show one taxol site per tubulin heterodimer, in agreement with the known stoichiometry of taxol in microtubules11. These studies indicate that taxol affects the interaction between protofilaments; to our knowledge, this is the first time that a ligand-binding site has been visualized in the tubulin molecule.

Original languageEnglish (US)
Pages (from-to)424-427
Number of pages4
JournalNature
Volume375
Issue number6530
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

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    Nogales, E., Grayer Wolf, S., Khan, I. A., Ludueña, R. F., & Downing, K. H. (1995). Structure of tubulin at 6.5 Å and location of the taxol-binding site. Nature, 375(6530), 424-427. https://doi.org/10.1038/375424a0