Proton-decoupled 15N and 31P solid-state NMR investigations of the Pf3 coat protein in oriented phospholipid bilayers.
The coat proteins of filamentous phage are first synthesized as transmembrane proteins and then assembled onto the extruding viral particles. We investigated the transmembrane conformation of the Pseudomonas aeruginosa Pf3 phage coat protein using proton-decoupled 15N and 31P solid-state NMR spectroscopy. The protein was either biochemically purified and uniformly labelled with 15N or synthesized chemically and labelled at specific sites. The proteins were then reconstituted into oriented phospholipid bilayers and the resulting samples analysed. The data suggest a model in which the protein adopts a tilted helix with an angle of approximately 30 degrees and an N-terminal 'swinging arm' at the membrane surface.
Aisenbrey
Christopher
C
Université Louis Pasteur/CNRS LC3-UMR71, Faculté de Chimie, Strasbourg, France.
Harzer
Ulrike
U
Bauer-Manz
Gabriele
G
Bär
Gerda
G
Chotimah
Irma N Husnal
IN
Bertani
Philippe
P
Sizun
Christina
C
Kuhn
Andreas
A
Bechinger
Burkhard
B
D036022:Capsid Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D002942:Circular Dichroism; D008051:Lipid Bilayers / Q000737:chemistry*; D008958:Models, Molecular; D009587:Nitrogen Isotopes / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D010759:Phosphorus Isotopes / Q000378:metabolism; D011487:Protein Conformation; D011522:Protons; D017105:Pseudomonas Phages / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D036022:Capsid Proteins; D008051:Lipid Bilayers; D009587:Nitrogen Isotopes; D010759:Phosphorus Isotopes; D011522:Protons; C084332:major coat protein, Pseudomonas phage Pf3