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Optimal molecular crowding accelerates group II intron folding and maximizes catalysis

Paudel, Bishnu P; Fiorini, Erica; Börner, Richard; Sigel, Roland K O; Rueda, David S (2018). Optimal molecular crowding accelerates group II intron folding and maximizes catalysis. Proceedings of the National Academy of Sciences of the United States of America, 115(47):11917-11922.

Abstract

Unlike in vivo conditions, group II intron ribozymes are known to require high magnesium(II) concentrations ([Mg<jats:sup>2+</jats:sup>]) and high temperatures (42 °C) for folding and catalysis in vitro. A possible explanation for this difference is the highly crowded cellular environment, which can be mimicked in vitro by macromolecular crowding agents. Here, we combined bulk activity assays and single-molecule Förster Resonance Energy Transfer (smFRET) to study the influence of polyethylene glycol (PEG) on catalysis and folding of the ribozyme. Our activity studies reveal that PEG reduces the [Mg<jats:sup>2+</jats:sup>] required, and we found an “optimum” [PEG] that yields maximum activity. smFRET experiments show that the most compact state population, the putative active state, increases with increasing [PEG]. Dynamic transitions between folded states also increase. Therefore, this study shows that optimal molecular crowding concentrations help the ribozyme not only to reach the native fold but also to increase its in vitro activity to approach that in physiological conditions.

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:07 Faculty of Science > Department of Chemistry
Dewey Decimal Classification:540 Chemistry
Scopus Subject Areas:Health Sciences > Multidisciplinary
Uncontrolled Keywords:Multidisciplinary
Language:English
Date:20 November 2018
Deposited On:09 Jan 2019 16:58
Last Modified:20 Oct 2024 01:37
Publisher:National Academy of Sciences
ISSN:0027-8424
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1073/pnas.1806685115
Project Information:
  • Funder: ERC
  • Grant ID:
  • Project Title: MIRNA 259092
  • Funder: UZH
  • Grant ID:
  • Project Title: FK-14-096
  • Funder: UZH
  • Grant ID:
  • Project Title: FK-15-095
  • Funder: COST Action CM1105
  • Grant ID:
  • Project Title:
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