Capturing Membrane Protein Ribosome Nascent Chain Complexes in a Native-like Environment for Co-translational Studies
Grant A. Pellowe, Heather E. Findlay, Karen Lee, Tim M Gemeinhardt, Laura R. Blackholly, Eamonn Reading, Paula J. Booth
Abstract
-generated membrane protein RNCs within their native lipid compositions. We reveal that a polytopic membrane protein can be successfully stalled at various stages during its synthesis and the resulting RNC extracted within either detergent micelles or diisobutylene-maleic acid co-polymer native nanodiscs. Our approaches offer tractable solutions for the structural and biophysical interrogation of nascent membrane proteins of specified lengths, as the elongating nascent chain emerges from the ribosome and inserts into its native lipid milieu.
Topics & Concepts
RibosomeNanodiscTransloconBiophysicsChemistryFolding (DSP implementation)Membrane proteinTranslation (biology)Protein biosynthesisProtein foldingCell biologyMembraneBiochemistryBiologyMessenger RNARNAGeneEngineeringElectrical engineeringRNA and protein synthesis mechanismsLipid Membrane Structure and BehaviorBacteriophages and microbial interactions