Light-coupled cryo-plunger for time-resolved cryo-EM

Nate Yoder, Farzad Jalali-Yazdi, Sigrid Noreng, Alexandra Houser, Isabelle Baconguis, Eric Gouaux

Research output: Contribution to journalArticlepeer-review

Abstract

Proteins are dynamic molecules that can undergo rapid conformational rearrangements in response to stimuli. These structural changes are often critical to protein function, and thus elucidating time-dependent conformational landscapes has been a long-standing goal of structural biology. To harness the power of single particle cryo-EM methods to enable ‘time-resolved’ structure determination, we have developed a light-coupled cryo-plunger that pairs flash-photolysis of caged ligands with rapid sample vitrification. The ‘flash-plunger’ consists of a high-power ultraviolet LED coupled with focusing optics and a motorized linear actuator, enabling the user to immobilize protein targets in vitreous ice within a programmable time window – as short as tens of milliseconds – after stimulus delivery. The flash-plunger is a simple, inexpensive and flexible tool to explore short-lived conformational states previously unobtainable by conventional sample preparation methods.

Original languageEnglish (US)
Article number107624
JournalJournal of Structural Biology
Volume212
Issue number3
DOIs
StatePublished - Dec 1 2020

Keywords

  • Cryo-EM
  • Cryo-electron microscopy
  • Manual cryo-plunger
  • Single particle
  • Time-resolved

ASJC Scopus subject areas

  • Structural Biology

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