tailieunhanh - Báo cáo Y học: NMR structure of the HIV-1 regulatory protein Vpr in H2O/trifluoroethanol Comparison with the Vpr N-terminal (1–51) and C-terminal (52–96) domains

The human immunodeficiency virus type 1, HIV-1, genome encodes a highly conserved regulatory gene product, Vpr (96 amino acids), which is incorporated into virions in quantities equivalent to those of the viral Gag protein. In infected cells, Vpr is believed to function during the early stages of HIV-1 replication (such as transcription of the proviral genome and migration of preintegration nuclear complex), blocks cells in G2 phase and triggers apoptosis. Vpr also plays a critical role in long-term AIDS disease by inducing viral infection in nondividing cells such as monocytes and macrophages. To gain deeper insight of the structure–function relationship. | Eur. J. Biochem. 269 3779-3788 2002 FEBS 2002 doi NMR structure of the HIV-1 regulatory protein Vpr in H2O trifluoroethanol Comparison with the Vpr N-terminal 1-51 and C-terminal 52-96 domains K. Wecker N. Morellet S. Bouaziz and B. P. Roques Departement de Pharmacochimie Moléculaire et Structurale INSERM U266 CNRS UMR 8600 UFR des Sciences Pharmaceutiques et Biologiques Paris France The human immunodeficiency virus type 1 HIV-1 genome encodes a highly conserved regulatory gene product Vpr 96 amino acids which is incorporated into virions in quantities equivalent to those of the viral Gag protein. In infected cells Vpr is believed to function during the early stages of HIV-1 replication such as transcription of the proviral genome and migration of preintegration nuclear complex blocks cells in G2 phase and triggers apoptosis. Vpr also plays a critical role in long-term AIDS disease by inducing viral infection in nondividing cells such as monocytes and macrophages. To gain deeper insight of the structure-function relationship of Vpr the intact protein residues 1-96 was synthezised. Its three-dimensional structure was analysed using circular dichroism and two-dimensional 1H- and 15N-NMR and refined by restrained molecular dynamics. In addition 15N relaxation parameters T1 T2 and heteronuclear 1H-15N NOEs were measured. The structure of the protein is characterized by a well-defined c turn 14-16 -a helix 17-33 -turn 34-36 followed by a a helix 40-48 -loop 49-54 -a helix 55-83 domain and ends with a very flexible C-terminal sequence. This structural determination of the whole intact Vpr molecule provide insights into the biological role played by this protein during the virus life cycle as such amphipathic helices are believed to be involved in protein-lipid bilayers protein-protein and or protein-nucleic acid interactions. Keywords Vpr NMR HIV-1 helix 3D structure. The genome of the human immunodeficiency virus type 1 HIV-1 the .