title: Motional restrictions of membrane proteins: a site-directed spin labeling study.
abstract: Site-directed mutagenesis was used to produce 27 single cysteine mutants of bacteriophage M13 major coat protein spanning the whole primary sequence of the protein. Single-cysteine mutants were labeled with nitroxide spin labels and incorporated into phospholipid bilayers with increasing acyl chain length. The SDSL is combined with ESR and CD spectroscopy. CD spectroscopy provided information about the overall protein conformation in different mismatching lipids. The spin label ESR spectra were analyzed in terms of a new spectral simulation approach based on hybrid evolutionary optimization and solution condensation. This method gives the residue-level free rotational space (i.e., the effective space within which the spin label can wobble) and the diffusion constant of the spin label attached to the protein. The results suggest that the coat protein has a large structural flexibility, which facilitates a stable protein-to-membrane association in lipid bilayers with various degrees of hydrophobic mismatch.
authors.affiliation: University of Ljubljana, Biotechnical Faculty, SI-1000 Ljubljana, Slovenia.
authors.forename: David
authors.initials: D
authors.lastname: Stopar
authors.forename: Janez
authors.initials: J
authors.lastname: Strancar
authors.forename: Ruud B
authors.initials: RB
authors.lastname: Spruijt
authors.forename: Marcus A
authors.initials: MA
authors.lastname: Hemminga
mesh_terms: D019943:Amino Acid Substitution; D036022:Capsid Proteins / Q000737:chemistry* / Q000648:ultrastructure*; D002942:Circular Dichroism / Q000379:methods; D003198:Computer Simulation; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D008956:Models, Chemical; D008958:Models, Molecular; D009038:Motion; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D013113:Spin Labels; D013329:Structure-Activity Relationship
chemical_list: D036022:Capsid Proteins; D013113:Spin Labels; C025668:coat protein, Bacteriophage M13