Solid-state NMR study of proteorhodopsin in the lipid environment: secondary structure and dynamics

Biochim Biophys Acta. 2009 Dec;1788(12):2563-74. doi: 10.1016/j.bbamem.2009.09.011. Epub 2009 Sep 30.

Abstract

Proteorhodopsins are typical retinal-binding light-driven proton pumps of heptahelical architecture widely distributed in marine and freshwater bacteria. Recently, we have shown that green proteorhodopsin (GPR) can be prepared in a lipid-bound state that gives well-resolved magic angle spinning (MAS) NMR spectra in samples with different patterns of reverse labelling. Here, we present 3D and 4D sequential chemical shift assignments identified through experiments conducted on a uniformly (13)C,(15)N-labelled sample. These experiments provided the assignments for 153 residues, with a particularly high density in the transmembrane regions ( approximately 74% of residues). The extent of assignments permitted a detailed examination of the secondary structure and dynamics in GPR. In particular, we present experimental evidence of mobility of the protein's termini and of the A-B, C-D, and F-G loops, the latter being possibly coupled to the GPR ion-transporting function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteria / chemistry
  • Bacteria / genetics
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Nuclear Magnetic Resonance, Biomolecular / methods
  • Protein Structure, Secondary / physiology
  • Rhodopsin / chemistry*
  • Rhodopsin / genetics
  • Rhodopsins, Microbial

Substances

  • Bacterial Proteins
  • Rhodopsins, Microbial
  • proteorhodopsin
  • Rhodopsin