Visualizing Specific Cross-Protomer Interactions in the Homo-Oligomeric Membrane Protein Proteorhodopsin by Dynamic-Nuclear-Polarization-Enhanced Solid-State NMR

作者: Jakob Maciejko , Michaela Mehler , Jagdeep Kaur , Tobias Lieblein , Nina Morgner

DOI: 10.1021/JACS.5B03606

关键词: Protein structureMembrane proteinLipid bilayerSolid-state nuclear magnetic resonancePeripheral membrane proteinPentamerChemistryProtomerCrystallographyProteorhodopsin

摘要: Membrane proteins often form oligomeric complexes within the lipid bilayer, but factors controlling their assembly are hard to predict and experimentally difficult determine. An understanding of protein–protein interactions bilayer is however required in order elucidate role oligomerization for functional mechanism stabilization. Here, we demonstrate pentameric, heptahelical membrane protein green proteorhodopsin that solid-state NMR could identify specific at protomer interfaces, if sensitivity enhanced by dynamic nuclear polarization. For this purpose, differently labeled protomers have been assembled into full pentamer complex embedded bilayer. We show proof concept one salt bridge determines formation pentamers or hexamers. Data supported laser-induced liquid bead ion desorption mass spectrometry blue native polyacrylamide gel electrophoresis analysis. The present...

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