Proteorhodopsin: characterisation of 2D crystals by electron microscopy and solid state NMR.
Proteorhodopsin (PR) a recent addition to retinal type 1 protein family, is a bacterial homologue of archaeal bacteriorhodopsin. It was found to high abundance in gamma-proteobacteria in the photic zone of the oceans and has been shown to act as a photoactive proton pump. It is therefore involved in the utilisation of light energy for energy production within the cell. Based on data from biodiversity screens, hundreds of variants were discovered worldwide, which are spectrally tuned to the available light at different locations in the sea. Here, we present a characterisation of 2D crystals of the green variant of proteorhodopsin by electron microscopy and solid state NMR. 2D crystal formation with hexagonal protein packing was observed under a very wide range of conditions indicating that PR might be also closely packed under native conditions. A low-resolution 2D projection map reveals a ring-shaped oligomeric assembly of PR. The protein state was analysed by 15N MAS NMR on lysine, tryptophan and methionine labelled samples. The chemical shift of the protonated Schiff base was almost identical to non-crystalline preparations. All residues could be cross-polarised in non-frozen samples. Lee-Goldberg cross-polarisation has been used to probe protein backbone mobility.
Shastri
Sarika
S
Institute of Biophysical Chemistry, Centre for Biomolecular Magnetic Resonance, J.W. Goethe University, Max von Laue Str. 9, D-60438 Frankfurt am Main, Germany.
Vonck
Janet
J
Pfleger
Nicole
N
Haase
Winfried
W
Kuehlbrandt
Werner
W
Glaubitz
Clemens
C
D003461:Crystallography / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008854:Microscopy, Electron / Q000379:methods*; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012243:Rhodopsin / Q000737:chemistry* / Q000648:ultrastructure; D025604:Rhodopsins, Microbial
D025604:Rhodopsins, Microbial; C413548:proteorhodopsin; D012243:Rhodopsin