title: 'Morphs' (MRFs): metal-reversible folding domains for differential IgG binding.
abstract: BACKGROUND
  Selective recognition and binding of IgG molecules is the basis for a host of immunological and affinity purification techniques. Capture of an IgG in these procedures relies chiefly on its interaction with one of a variety of reengineered bacterial receptors which bind to the Fc region of IgG molecules with very high affinity. While this interaction is extremely efficient in trapping IgG molecules, the tight interaction between the binding partners often requires denaturing conditions for disruption of the complex, which can adversely affect the yield of purified IgG and also limit the lifetime of the receptor matrix. An effective receptor/IgG binding system which could be modulated by less extreme conditions is of considerable general interest.
  
  
  RESULTS
  We describe the properties of a series of modified Fc receptor domains which are competent to bind IgG with high affinity but which can be reversibly unfolded upon addition of modest amounts of transition metal ions. Data are presented demonstrating loss of the secondary structural content of the domains as a function of increasing metal concentration, with a concomitant decrease in IgG binding affinity. Variants of the Fc receptor differing at a single amino acid position display increased sensitivity to metal-induced unfolding, while retaining comparable IgG binding ability in the absence of metal.
  
  
  CONCLUSIONS
  The interaction of this series of Fc receptors with metal ions abolishes IgG binding, but removal of metal ions allows refolding of the domains with restoration of the IgG binding function. Examples of the utility and potential applications of these metal-modulated IgG binding domains are discussed.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, 266 Whitney Avenue, New Haven, CT 06520, USA. smarino@bioc.unizh.ch
authors.forename: S F
authors.initials: SF
authors.lastname: Marino
authors.forename: D
authors.initials: D
authors.lastname: Shechner
authors.forename: L
authors.initials: L
authors.lastname: Regan
mesh_terms: D001667:Binding, Competitive; D002942:Circular Dichroism; D007074:Immunoglobulin G / Q000378:metabolism*; D008670:Metals; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011961:Receptors, Fc / Q000737:chemistry* / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D007074:Immunoglobulin G; D008670:Metals; D011961:Receptors, Fc