Dynamic Nuclear Polarization Illuminates Key Protein-Lipid Interactions in the Native Bacterial Cell Envelope. Biochemistry 2023 Aug 01;62(15):2252-2256
Date
07/17/2023Pubmed ID
37459255Pubmed Central ID
PMC11019665DOI
10.1021/acs.biochem.3c00262Scopus ID
2-s2.0-85166390720 (requires institutional sign-in at Scopus site) 1 CitationAbstract
Elucidating the structure and interactions of proteins in native environments is a fundamental goal of structural biology. Nuclear magnetic resonance (NMR) spectroscopy is well suited for this task but often suffers from low sensitivity, especially in complex biological settings. Here, we use a sensitivity-enhancement technique called dynamic nuclear polarization (DNP) to overcome this challenge. We apply DNP to capture the membrane interactions of the outer membrane protein Ail, a key component of the host invasion pathway of Yersinia pestis. We show that the DNP-enhanced NMR spectra of Ail in native bacterial cell envelopes are well resolved and enriched in correlations that are invisible in conventional solid-state NMR experiments. Furthermore, we demonstrate the ability of DNP to capture elusive interactions between the protein and the surrounding lipopolysaccharide layer. Our results support a model where the extracellular loop arginine residues remodel the membrane environment, a process that is crucial for host invasion and pathogenesis.
Author List
Kent JE, Ackermann BE, Debelouchina GT, Marassi FMAuthor
Francesca M. Marassi PhD Chair, Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Cell MembraneCell Wall
Lipids
Magnetic Resonance Spectroscopy
Membrane Proteins
Nuclear Magnetic Resonance, Biomolecular
