title: Analyses of the stability and function of three surface mutants (R82C, K69H, and L32R) of the gene V protein from Ff phage by X-ray crystallography.
abstract: The high-resolution crystal structure of the gene V protein (GVP) from the Ff filamentous phages (M13, fl, fd) has been solved recently for the wild-type and two surface mutant (Y41F and Y41H) proteins, leading to a plausible model for the polymeric GVP-ssDNA complex (Guan Y, Zhang H, Wang AHJ, 1995, Protein Sci 4:187-197). The model of the complex shows extensive contacts between neighboring dimer GVPs involving electrostatic interactions between the K69 from one and the D79 and R82 from the next dimer. In addition, hydrophobic interactions between the amino acids L32 and L44 from one and G23 from the next dimer also contribute to the dimer-dimer interactions. Mutations at the L32, K69, and R82 amino acid sites generally destabilize the protein and many of these affect the function of the phage. We have studied the structural effects of three mutant proteins involving those sites, i.e., L32R, K69H, and R82C, by X-ray crystallographic analysis at 2.0 A resolution. In L32R GVP, the structural perturbation is localized, whereas in K69H and R82C GVPs, some long-range effects are also detected in addition to the local perturbation. We have interpreted the protein stability and the functional properties associated with those mutations in terms of the observed structural perturbations.
authors.affiliation: Department of Cell & Structural Biology, University of Illinois at Urbana-Champaign 61801, USA.
authors.forename: S
authors.initials: S
authors.lastname: Su
authors.forename: Y G
authors.initials: YG
authors.lastname: Gao
authors.forename: H
authors.initials: H
authors.lastname: Zhang
authors.forename: T C
authors.initials: TC
authors.lastname: Terwilliger
authors.forename: A H
authors.initials: AH
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000737:chemistry*; D018360:Crystallography, X-Ray; D008969:Molecular Sequence Data; D009154:Mutation; D055672:Static Electricity; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D014764:Viral Proteins; C069558:gene V protein, Enterobacteria phage f1