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Tilt-series for the electron cryotomogram of Vibrio cholerae O395 N1

ABSTRACT:

SUBMITTER: Yi-Wei Chang

PROVIDER: EMPIAR-10110 | biostudies-other |

REPOSITORIES: biostudies-other

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Architecture of the Vibrio cholerae toxin-coregulated pilus machine revealed by electron cryotomography.

Chang Yi-Wei YW Kjær Andreas A Ortega Davi R DR Kovacikova Gabriela G Sutherland John A JA Rettberg Lee A LA Taylor Ronald K RK Jensen Grant J GJ

Nature microbiology 20170206

Type IV pili (T4P) are filamentous appendages found on many Bacteria and Archaea. They are helical fibres of pilin proteins assembled by a multi-component macromolecular machine we call the basal body. Based on pilin features, T4P are classified into type IVa pili (T4aP) and type IVb pili (T4bP)1,2. T4aP are more widespread and are involved in cell motility3, DNA transfer4, host predation5 and electron transfer6. T4bP are less prevalent and ...[more]

PMID: 28165453

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Rapid tilt-series acquisition for electron cryotomography.

Project description:Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a tilt-series in seconds, but with loss of details finer than ?4?nm. We also demonstrate a fast-incremental method that can record a tilt-series several-fold faster than current methods and with similar resolution. We characterize the utility of both methods in real biological electron cryotomography workflows. We identify opportunities for further improvements in hardware and software and speculate on the impact such advances could have on structural biology.

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