title: Probing the structure of the Ff bacteriophage major coat protein transmembrane helix dimer by solution NMR.
abstract: The transmembrane (TM) segment of the major coat protein from Ff bacteriophage has been extensively studied as an example of dimerization in detergent and lipid bilayer systems. However, almost all the information regarding this interaction has been gained through mutagenesis studies, with little direct structural information being available. To this end solution NMR has the potential to provide new insights into structure of the dimer. In order to evaluate the utility of this approach we have studied a selectively 15N-labeled peptide containing the TM segment of MCP (MCPTM) by solution NMR. This peptide was found to give rise to detergent concentration-dependent spectra that were assigned to monomeric and dimeric forms. The standard free energy of this interaction in SDS was estimated from these spectra and found to be consistent with weak but specific dimerization. In addition, similar spectra could be obtained in beta-octyl glucoside with intermolecular paramagnetic relaxation experiments demonstrating a parallel arrangement of TM helices in the dimer. In both detergents backbone chemical shift differences between monomeric and dimeric forms of MCPTM showed that the largest changes occur around its GXXXG motif. The resulting structural model is consistent with observations made for MCP mutants previously characterized in biological membranes, opening the door to detailed structural characterization of this form of MCP. These results also have general implications for the study of weakly interacting TM segments by solution NMR since the use of similar sample conditions should allow structural data to be accessed for oligomeric states from a wide range systems that undergo biologically relevant but weak associations in the membrane.
authors.affiliation: Department of Chemistry, University of Ottawa, 10 Marie Curie, Ottawa, ON, K1N 6N5 Canada.
authors.forename: Yanqiu
authors.initials: Y
authors.lastname: Wu
authors.forename: Steve C C
authors.initials: SC
authors.lastname: Shih
authors.forename: Natalie K
authors.initials: NK
authors.lastname: Goto
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000378:metabolism*; D036022:Capsid Proteins / Q000737:chemistry*; D002942:Circular Dichroism; D019281:Dimerization; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008823:Micelles; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012996:Solutions
chemical_list: D036022:Capsid Proteins; D008823:Micelles; D012996:Solutions