Correlation of membrane protein conformational and functional dynamics

Raghavendar Reddy Sanganna Gari, Joel José Montalvo‐Acosta, George R. Heath, Yining Jiang, Xiaolong Gao, Crina M. Nimigean, Christophe Chipot () and Simon Scheuring ()
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Raghavendar Reddy Sanganna Gari: Weill Cornell Medicine, Department of Anesthesiology
Joel José Montalvo‐Acosta: Laboratoire International Associé CNRS and University of Illinois at Urbana-Champaign, Université de Lorraine
George R. Heath: Weill Cornell Medicine, Department of Anesthesiology
Yining Jiang: Weill Cornell Medicine, Department of Physiology and Biophysics
Xiaolong Gao: Weill Cornell Medicine, Department of Anesthesiology
Crina M. Nimigean: Weill Cornell Medicine, Department of Anesthesiology
Christophe Chipot: Laboratoire International Associé CNRS and University of Illinois at Urbana-Champaign, Université de Lorraine
Simon Scheuring: Weill Cornell Medicine, Department of Anesthesiology

Nature Communications, 2021, vol. 12, issue 1, 1-11

Abstract: Abstract Conformational changes in ion channels lead to gating of an ion-conductive pore. Ion flux has been measured with high temporal resolution by single-channel electrophysiology for decades. However, correlation between functional and conformational dynamics remained difficult, lacking experimental techniques to monitor sub-millisecond conformational changes. Here, we use the outer membrane protein G (OmpG) as a model system where loop-6 opens and closes the β-barrel pore like a lid in a pH-dependent manner. Functionally, single-channel electrophysiology shows that while closed states are favored at acidic pH and open states are favored at physiological pH, both states coexist and rapidly interchange in all conditions. Using HS-AFM height spectroscopy (HS-AFM-HS), we monitor sub-millisecond loop-6 conformational dynamics, and compare them to the functional dynamics from single-channel recordings, while MD simulations provide atomistic details and energy landscapes of the pH-dependent loop-6 fluctuations. HS-AFM-HS offers new opportunities to analyze conformational dynamics at timescales of domain and loop fluctuations.

Date: 2021
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DOI: 10.1038/s41467-021-24660-1

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