tailieunhanh - Báo cáo khoa học: 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 transmem-brane proteins and then assembled onto the extruding viral particles. We investigated the transmembrane conformation of the Pseudomonas aerugi-nosa Pf3 phage coat protein using proton-decoupled 15 N and 31 P solid-state NMR spectroscopy. | ềFEBS Journal Proton-decoupled 15N and 31P solid-state NMR investigations of the Pf3 coat protein in oriented phospholipid bilayers Christopher Aisenbrey1 2 Ulrike Harzer2 Gabriele Bauer-Manz3 Gerda Bar3 Irma N. Husnal Chotimah1 Philippe Bertani1 Christina Sizun2 Andreas Kuhn3 and Burkhard Bechinger1 2 1 Université Louis Pasteur CNRS LC3-UMR71 Faculte de Chimie Strasbourg France 2 Max-Planck-Institut fur Biochemie Martinsried Germany 3 Institute of Microbiology and Molecular Biology University of Hohenheim Stuttgart Germany Keywords oriented lipid bilayer amphipathic a helix membrane protein structure protein dynamics mosaic spread Correspondence B. Bechinger Faculte de chimie Institut le Bel 4 rue Blaise Pascal 67070 Strasbourg France Fax 33 3 90 24 51 51 Tel 33 3 90 24 51 50 E-mail bechinger@ 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 w 30 and an N-terminal swinging arm at the membrane surface. These authors contributed equally to this work. Received 20 July 2005 revised 19 December 2005 accepted 21 December 2005 doi During the reproductive life cycle of filamentous phage the major coat protein adopts many different conformational states. After synthesis in the bacterial cytoplasm it is readily bound to the membrane surface and inserted by the membrane insertase YidC in a Secindependent fashion 1 2 . In contrast to Escherichia .
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