title: Packing of coat protein amphipathic and transmembrane helices in filamentous bacteriophage M13: role of small residues in protein oligomerization.
abstract: Filamentous bacteriophage M13, an important cloning and phage display vector, is encapsulated by ca 2700 copies of its 50-residue major coat protein (gene 8). This protein occurs as a membrane protein while stably inserted into its E. coli host inner membrane, and as a coat protein upon assembly and packing onto phage DNA in the lipid-free virion. To examine the specific protein-protein interactions underlying these processes, we used a combination of randomized and saturation mutagenesis of the entire gene 8 to assess the susceptibility of each position to mutation. In the resulting library of ca 100 viable M13 mutants, "small" residues (Ala,Gly,Ser), which constitute the non-polar face of the N-terminal amphipathic helical segment, and a face of the hydrophobic (effective transmembrane) helical segment, were found to be highly conserved. These results support a model in which coat protein packing is stabilized by the presence within each protein subunit of two "oligomerization segments", i.e. specific helical regions with faces rich in small residues which function to promote the close approach of alpha-helices.
authors.affiliation: Division of Biochemistry Research, Research Institute Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: K A
authors.initials: KA
authors.lastname: Williams
authors.forename: M
authors.initials: M
authors.lastname: Glibowicka
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.forename: A R
authors.initials: AR
authors.lastname: Khan
authors.forename: Y M
authors.initials: YM
authors.lastname: Chen
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: D A
authors.initials: DA
authors.lastname: Marvin
authors.forename: C M
authors.initials: CM
authors.lastname: Deber
mesh_terms: D000595:Amino Acid Sequence; D017104:Bacteriophage M13 / Q000648:ultrastructure*; D001483:Base Sequence; D002213:Capsid / Q000648:ultrastructure*; D017931:DNA Primers / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009024:Morphogenesis; D016297:Mutagenesis, Site-Directed; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
chemical_list: D017931:DNA Primers