title: Membrane partitioning of the pore-forming domain of colicin A. Role of the hydrophobic helical hairpin.
abstract: The colicins are bacteriocins that target Escherichia coli and kill bacterial cells through different mechanisms. Colicin A forms ion channels in the inner membranes of nonimmune bacteria. This activity resides exclusively in its C-terminal fragment (residues 387-592). The soluble free form of this domain is a 10 α-helix bundle. The hydrophobic helical hairpin, H8-H9, is buried inside the structure and shielded by eight amphipathic surface helices. The interaction of the C-terminal colicin A domain and several chimeric variants with lipidic vesicles was examined here by isothermal titration calorimetry. In the mutant constructions, natural sequences of the hydrophobic helices H8 and H9 were either removed or substituted by polyalanine or polyleucine. All the constructions fully associated with DOPG liposomes including the mutant that lacked helices H8 and H9, indicating that amphipathic rather than hydrophobic helices were the major determinants of the exothermic binding reactions. Alanine is not specially favored in the lipid-bound form; the chimeric construct with polyalanine produced lower enthalpy gain. On the other hand, the large negative heat capacities associated with partitioning, a characteristic feature of the hydrophobic effect, were found to be dependent on the sequence hydrophobicity of helices H8 and H9.
authors.affiliation: Unidad de Biofísica (CSIC, UPV/EHU), Barrio Sarriena s/n, Leioa, Spain.
authors.forename: Ivan L
authors.initials: IL
authors.lastname: Bermejo
authors.forename: Cristina
authors.initials: C
authors.lastname: Arnulphi
authors.forename: Alain
authors.initials: A
authors.lastname: Ibáñez de Opakua
authors.forename: Marián
authors.initials: M
authors.lastname: Alonso-Mariño
authors.forename: Félix M
authors.initials: FM
authors.lastname: Goñi
authors.forename: Ana R
authors.initials: AR
authors.lastname: Viguera
mesh_terms: D019943:Amino Acid Substitution; D002462:Cell Membrane / Q000378:metabolism*; D003087:Colicins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D010715:Phosphatidylglycerols / Q000378:metabolism; D016062:Porosity; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013816:Thermodynamics
chemical_list: D003087:Colicins; D010715:Phosphatidylglycerols; C051388:1,2-dioleoyl-sn-glycero-3-phosphoglycerol
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