Rxivist logo

Rxivist combines preprints from bioRxiv with data from Twitter to help you find the papers being discussed in your field. Currently indexing 73,401 bioRxiv papers from 319,519 authors.

The resolution revolution in cryoEM requires new sample preparation procedures: A rapid pipeline to high resolution maps of yeast FAS

By Mirko Joppe, Edoardo D’Imprima, Nina Salustros, Karthik S. Paithankar, Janet Vonck, Martin Grininger, Werner Kühlbrandt

Posted 04 Nov 2019
bioRxiv DOI: 10.1101/829176 (published DOI: 10.1107/S2052252519017366)

Single-particle electron cryo-microscopy (cryoEM) has undergone a "resolution revolution" that makes it possible to characterize megadalton (MDa) complexes at atomic resolution without crystals. To fully exploit the new opportunities in molecular microscopy, new procedures for the cloning, expression and purification of macromolecular complexes need to be explored. Macromolecular assemblies are often unstable, and invasive construct design or inadequate purification conditions or sample preparation methods can result in disassembly or denaturation. The structure of the 2.6 MDa yeast fatty acid synthase (FAS) has been studied by electron microscopy since the 1960s. We report a new, streamlined protocol for the rapid production of purified yeast FAS for structure determination by high-resolution cryoEM. Together with a companion protocol for preparing cryoEM specimens on a hydrophilized graphene layer, our new protocol has yielded a 3.1 Å map of yeast FAS from 15,000 automatically picked particles within a day. The high map quality enabled us to build a complete atomic model of an intact fungal FAS.

Download data

  • Downloaded 539 times
  • Download rankings, all-time:
    • Site-wide: 21,710 out of 73,413
    • In biochemistry: 507 out of 2,231
  • Year to date:
    • Site-wide: 20,984 out of 73,413
  • Since beginning of last month:
    • Site-wide: 20,984 out of 73,413

Altmetric data


Downloads over time

Distribution of downloads per paper, site-wide


PanLingua

Sign up for the Rxivist weekly newsletter! (Click here for more details.)


News