Journey of bacteriophage M13 major coat protein.ppt

Journey of bacteriophage M13 major coat protein Replication cycle of the bacteriophage M13 Structure of the major coat protein Topology of the major coat protein in the lipid bilayer Anchoring of the major coat protein in the lipid bilayer Schematic representation of bacteriophage M13 replication cycle Model of the of bacteriophage M13 protein coat Nearest-neighbour interactions between protein subunits in the phage particle Major coat protein structural domains in lipid bilayer X-ray and NMR limitations for membrane proteins M13 Coat Protein with Labels Attached Sensitivity of spin labels for different lipids Spin label relaxation times in DOPC bilayers Relative quenching efficiency by oxygen and Ni2+ Dependence of spin labeled DOPC acyl chains on the relaxation enhancement by oxygen and Ni2+ Location of the protein in the lipid bilayer as determined by fluorescence labeling AEDANS lmax of the A7C protein mutants in lipid bilayers AEDANS lmax in DOPC lipid bilayers Tryptophane lmax in DOPC lipid bilayers Bacteriophage M13 major coat protein anchors in the lipid bilayer Model of the major coat protein disassembly and assembly * David Stopar University of Ljubljana Slovenia Interactions in macromolecular assemblies Journey of bacteriophage M13 major coat protein M13 E.coli F-pilus DNA gp 5 gp 8 Gene 9 protein Gene 6 protein Gene 3 protein Gene 8 protein Circular ssDNA Distal end Proximal end Gene 7 protein Schematic structure of bacteriophage M13 filament k=11 k=11 k=6 k=6 k=0 k=0 Replication cycle of the bacteriophage M13 Structure of the major coat protein Topology of the major coat protein in the lipid bilayer Anchoring of the major coat protein in the lipid bilayer Journey of bacteriophage M13 major coat protein 1-5 N-terminus 6-16 amphiphatic helix 17-23 loop 24-46 transmembrane helix 47-50 C-terminus 1 50 Amino acid residue number X-ray: “no” crystals of membrane proteins NMR: membranes are anisotropic systems high-resolution NMR limited to