Cryo-electron microscopy of GDP-tubulin rings

William V. Nicholson, Minou Lee, Kenneth H. Downing, Eva Nogales

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Rings of guanosine diphosphate (GDP)-tubulin formed in the presence of divalent cations have been studied using conventional negative stain and cryo-electron microscopy. The structure of such rings resembles that of depolymerizing microtubule ends and corresponds to an “unconstrained” conformation of tubulin in its GDP state. The use of cryo-techniques has allowed us to image the ring polymers free from dehydration and flattening artifacts. Preparations of frozenhydrated GDP-tubulin rings are generally heterogeneous and contain a mixture of double, triple, and incomplete rings, as well as spirals and some rare single rings. Images of different polymer types can be identified and classified into groups that are then amenable for averaging and single particle reconstruction methods. Identifying the differences in tubulin structure, between straight and curve protofilaments, will be important to understand the molecular bases of dynamic instability in microtubules.
Original languageEnglish
Pages (from-to)175-183
Number of pages9
JournalCell Biochem Biophys
Volume31
Issue number2
DOIs
Publication statusPublished - Jun 1999

Fingerprint

Guanosine
Diphosphates
Tubulin
Electron microscopy
Polymers
Divalent Cations
Dehydration
Conformations
Coloring Agents

Keywords

  • Tubulin
  • microtubules
  • GDP
  • depolymerization
  • cryo-electron microscopy
  • single particles

Cite this

Nicholson, W. V., Lee, M., Downing, K. H., & Nogales, E. (1999). Cryo-electron microscopy of GDP-tubulin rings. Cell Biochem Biophys, 31(2), 175-183. https://doi.org/10.1007/bf02738171
Nicholson, William V. ; Lee, Minou ; Downing, Kenneth H. ; Nogales, Eva. / Cryo-electron microscopy of GDP-tubulin rings. In: Cell Biochem Biophys. 1999 ; Vol. 31, No. 2. pp. 175-183.
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Nicholson, WV, Lee, M, Downing, KH & Nogales, E 1999, 'Cryo-electron microscopy of GDP-tubulin rings', Cell Biochem Biophys, vol. 31, no. 2, pp. 175-183. https://doi.org/10.1007/bf02738171

Cryo-electron microscopy of GDP-tubulin rings. / Nicholson, William V.; Lee, Minou; Downing, Kenneth H.; Nogales, Eva.

In: Cell Biochem Biophys, Vol. 31, No. 2, 06.1999, p. 175-183.

Research output: Contribution to journalArticle

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T1 - Cryo-electron microscopy of GDP-tubulin rings

AU - Nicholson, William V.

AU - Lee, Minou

AU - Downing, Kenneth H.

AU - Nogales, Eva

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AB - Rings of guanosine diphosphate (GDP)-tubulin formed in the presence of divalent cations have been studied using conventional negative stain and cryo-electron microscopy. The structure of such rings resembles that of depolymerizing microtubule ends and corresponds to an “unconstrained” conformation of tubulin in its GDP state. The use of cryo-techniques has allowed us to image the ring polymers free from dehydration and flattening artifacts. Preparations of frozenhydrated GDP-tubulin rings are generally heterogeneous and contain a mixture of double, triple, and incomplete rings, as well as spirals and some rare single rings. Images of different polymer types can be identified and classified into groups that are then amenable for averaging and single particle reconstruction methods. Identifying the differences in tubulin structure, between straight and curve protofilaments, will be important to understand the molecular bases of dynamic instability in microtubules.

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Nicholson WV, Lee M, Downing KH, Nogales E. Cryo-electron microscopy of GDP-tubulin rings. Cell Biochem Biophys. 1999 Jun;31(2):175-183. https://doi.org/10.1007/bf02738171