title: Control of electrostatic interactions between F-actin and genetically modified lysozyme in aqueous media.
abstract: The aim for deterministic control of the interactions between macroions in aqueous media has motivated widespread experimental and theoretical work. Although it has been well established that like-charged macromolecules can aggregate under the influence of oppositely charged condensing agents, the specific conditions for the stability of such aggregates can only be determined empirically. We examine these conditions, which involve an interplay of electrostatic and osmotic effects, by using a well defined model system composed of F-actin, an anionic rod-like polyelectrolyte, and lysozyme, a cationic globular protein with a charge that can be genetically modified. The structure and stability of actin-lysozyme complexes for different lysozyme charge mutants and salt concentrations are examined by using synchrotron x-ray scattering and molecular dynamics simulations. We provide evidence that supports a structural transition from columnar arrangements of F-actin held together by arrays of lysozyme at the threefold interstitial sites of the actin sublattice to marginally stable complexes in which lysozyme resides at twofold bridging sites between actin. The reduced stability arises from strongly reduced partitioning of salt between the complex and the surrounding solution. Changes in the stability of actin-lysozyme complexes are of biomedical interest because their formation has been reported to contribute to the persistence of airway infections in cystic fibrosis by sequestering antimicrobials such as lysozyme. We present x-ray microscopy results that argue for the existence of actin-lysozyme complexes in cystic fibrosis sputum and demonstrate that, for a wide range of salt conditions, charge-reduced lysozyme is not sequestered in ordered complexes while retaining its bacterial killing activity.
authors.affiliation: Department of Materials Science and Engineering, The Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801-2920, USA.
authors.forename: Lori K
authors.initials: LK
authors.lastname: Sanders
authors.forename: Wujing
authors.initials: W
authors.lastname: Xian
authors.forename: Camilo
authors.initials: C
authors.lastname: Guáqueta
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Strohman
authors.forename: Chuck R
authors.initials: CR
authors.lastname: Vrasich
authors.forename: Erik
authors.initials: E
authors.lastname: Luijten
authors.forename: Gerard C L
authors.initials: GC
authors.lastname: Wong
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology; D017122:Bacteriophage T4 / Q000201:enzymology / Q000235:genetics; D001665:Binding Sites; D004795:Enzyme Stability; D066298:In Vitro Techniques; D008958:Models, Molecular; D046912:Multiprotein Complexes; D009113:Muramidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D016297:Mutagenesis, Site-Directed; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D054712:Salinity; D053838:Scattering, Small Angle; D055672:Static Electricity; D013816:Thermodynamics; D014867:Water; D014961:X-Ray Diffraction
chemical_list: D000199:Actins; D000900:Anti-Bacterial Agents; D046912:Multiprotein Complexes; D011994:Recombinant Proteins; D014867:Water; D009113:Muramidase